Adam Sperling, MD, PhD

All Publications

• PPM1D modulates hematopoietic cell fitness and response to DNA damage and is a therapeutic target in myeloid malignancy BLOOD Miller, P. G., Sperling, A. S., Mayerhofer, C., McConkey, M. E., Ellegast, J. M., Da Silva, C., Cohen, D. N., Wang, C., Sharda, A., Yan, N., Saha, S., Schluter, C., Schechter, I., Slabicki, M., Sandoval, B., Kahn, J., Boettcher, S., Gibson, C. J., Scadden, D. T., Stegmaier, K., Bhatt, S., Lindsley, R., Ebert, B. L. 2023; 142 (24): 2079-2091

  Abstract

  PPM1D encodes a phosphatase that is recurrently activated across cancer, most notably in therapy-related myeloid neoplasms. However, the function of PPM1D in hematopoiesis and its contribution to tumor cell growth remain incompletely understood. Using conditional mouse models, we uncover a central role for Ppm1d in hematopoiesis and validate its potential as a therapeutic target. We find that Ppm1d regulates the competitive fitness and self-renewal of hematopoietic stem cells (HSCs) with and without exogenous genotoxic stresses. We also show that although Ppm1d activation confers cellular resistance to cytotoxic therapy, it does so to a lesser degree than p53 loss, informing the clonal competition phenotypes often observed in human studies. Notably, loss of Ppm1d sensitizes leukemias to cytotoxic therapies in vitro and in vivo, even in the absence of a Ppm1d mutation. Vulnerability to PPM1D inhibition is observed across many cancer types and dependent on p53 activity. Importantly, organism-wide loss of Ppm1d in adult mice is well tolerated, supporting the tolerability of pharmacologically targeting PPM1D. Our data link PPM1D gain-of-function mutations to the clonal expansion of HSCs, inform human genetic observations, and support the therapeutic targeting of PPM1D in cancer.

  View details for DOI 10.1182/blood.2023020331

  View details for Web of Science ID 001146280500001

  View details for PubMedID 37595362

  View details for PubMedCentralID PMC10733824

• Clinical Outcomes and Evolution of Clonal Hematopoiesis in Patients with Newly Diagnosed Multiple Myeloma CANCER RESEARCH COMMUNICATIONS Mouhieddine, T. H., Nzerem, C., Redd, R., Dunford, A., Leventhal, M., Sklavenitis-Pistofidis, R., Tahri, S., El-Khoury, H., Steensma, D. P., Ebert, B. L., Soiffer, R. J., Keats, J. J., Mehr, S., Auclair, D., Ghobrial, I. M., Sperling, A. S., Stewart, C., Getz, G. 2023; 3 (12): 2560-2571

  Abstract

  Clonal hematopoiesis (CH) at time of autologous stem cell transplant (ASCT) has been shown to be associated with decreased overall survival (OS) and progression-free survival (PFS) in patients with multiple myeloma not receiving immunomodulatory drugs (IMiD). However, the significance of CH in newly diagnosed patients, including transplant ineligible patients, and its effect on clonal evolution during multiple myeloma therapy in the era of novel agents, has not been well studied. Using our new algorithm to differentiate tumor and germline mutations from CH, we detected CH in approximately 10% of 986 patients with multiple myeloma from the Clinical Outcomes in MM to Personal Assessment of Genetic Profile (CoMMpass) cohort (40/529 transplanted and 59/457 non-transplanted patients). CH was associated with increased age, risk of recurrent bacterial infections and cardiovascular disease. CH at time of multiple myeloma diagnosis was not associated with inferior OS or PFS regardless of undergoing ASCT, and all patients benefited from IMiD-based therapies, irrespective of the presence of CH. Serial sampling of 52 patients revealed the emergence of CH over a median of 3 years of treatment, increasing its prevalence to 25%, mostly with DNMT3A mutations.Using our algorithm to differentiate tumor and germline mutations from CH mutations, we detected CH in approximately 10% of patients with newly diagnosed myeloma, including both transplant eligible and ineligible patients. Receiving IMiDs improved outcomes irrespective of CH status, but the prevalence of CH significantly rose throughout myeloma-directed therapy.

  View details for DOI 10.1158/2767-9764.CRC-23-0093

  View details for Web of Science ID 001178977700012

  View details for PubMedID 38019104

  View details for PubMedCentralID PMC10730502

• Degradation of GSPT1 causes TP53-independent cell death in leukemia while sparing normal hematopoietic stem cells JOURNAL OF CLINICAL INVESTIGATION Sellar, R. S., Sperling, A. S., Slabicki, M., Gasser, J. A., McConkey, M. E., Donovan, K. A., Mageed, N., Adams, D. N., Zou, C., Miller, P. G., Dutta, R. K., Boettcher, S., Lin, A. E., Sandoval, B., Barrios, V., Kovalcik, V., Koeppel, J., Henderson, E. K., Fink, E. C., Yang, L., Chan, A., Pokharel, S., Bergstrom, E. J., Burt, R., Udeshi, N. D., Carr, S. A., Fischer, E. S., Chen, C., Ebert, B. L. 2022; 132 (16)

  Abstract

  Targeted protein degradation is a rapidly advancing and expanding therapeutic approach. Drugs that degrade GSPT1 via the CRL4CRBN ubiquitin ligase are a new class of cancer therapy in active clinical development with evidence of activity against acute myeloid leukemia in early-phase trials. However, other than activation of the integrated stress response, the downstream effects of GSPT1 degradation leading to cell death are largely undefined, and no murine models are available to study these agents. We identified the domains of GSPT1 essential for cell survival and show that GSPT1 degradation leads to impaired translation termination, activation of the integrated stress response pathway, and TP53-independent cell death. CRISPR/Cas9 screens implicated decreased translation initiation as protective following GSPT1 degradation, suggesting that cells with higher levels of translation are more susceptible to the effects of GSPT1 degradation. We defined 2 Crbn amino acids that prevent Gspt1 degradation in mice, generated a knockin mouse with alteration of these residues, and demonstrated the efficacy of GSPT1-degrading drugs in vivo with relative sparing of numbers and function of long-term hematopoietic stem cells. Our results provide a mechanistic basis for the use of GSPT1 degraders for the treatment of cancer, including TP53-mutant acute myeloid leukemia.

  View details for DOI 10.1172/JCI153514

  View details for Web of Science ID 000856124500002

  View details for PubMedID 35763353

  View details for PubMedCentralID PMC9374383

• Clonal hematopoiesis is associated with increased risk of progression of asymptomatic Waldenstrom macroglobulinemia BLOOD ADVANCES Tahri, S., Mouhieddine, T. H., Redd, R., Lampe, L., Nilsson, K., El-Khoury, H., Su, N., Nassar, A. H., Adib, E., Bindra, G., Abou Alaiwi, S., Trippa, L., Steensma, D. P., Castillo, J. J., Treon, S. P., Ghobrial, I. M., Sperling, A. S. 2022; 6 (7): 2230-2235

  Abstract

  Clonal hematopoiesis (CH) is associated with adverse outcomes in patients with non-Hodgkin lymphoma (NHL) and multiple myeloma undergoing autologous stem cell transplantation. Still, its implications for patients with indolent NHL have not been well studied. We report the prevalence of CH in patients with Waldenström macroglobulinemia (WM) and its association with clinical outcomes. To unambiguously differentiate CH mutations from those in the WM clone, CH was defined by the presence of somatic mutations in DNMT3A, TET2, or ASXL1 (DTA) and was detected in 14% of 587 patients with IgM monoclonal gammopathy of undetermined significance (MGUS), smoldering WM (SWM) or WM. The presence and size of DTA clones were associated with older age. Patients with CH had an increased risk of progression from MGUS or SWM to WM, but not worse overall survival in this cohort. These findings further illuminate the clinical effects of CH in patients with indolent NHL such as WM.

  View details for DOI 10.1182/bloodadvances.2021004926

  View details for Web of Science ID 000792426300006

  View details for PubMedID 34847227

  View details for PubMedCentralID PMC9006277

• Characteristics and Outcomes of Eosinophilic Fasciitis-Associated Monoclonal Gammopathy JAMA DERMATOLOGY Mazori, D. R., Kassamali, B., Min, M. S., Schaefer, M., LaChance, A. H., Sperling, A. S., Vleugels, R. 2021; 157 (12): 1508-1509

  Abstract

  This cross-sectional study analyzes the characteristics and outcomes of eosinophilic fasciitis–associated monoclonal gammopathy.

  View details for DOI 10.1001/jamadermatol.2021.4302

  View details for Web of Science ID 000712536500003

  View details for PubMedID 34705045

  View details for PubMedCentralID PMC8552112

• Facts and Hopes in Multiple Myeloma Immunotherapy CLINICAL CANCER RESEARCH Sperling, A. S., Anderson, K. C. 2021; 27 (16): 4468-4477

  Abstract

  Among the hallmarks of cancer is the ability of neoplastic cells to evade and suppress immune surveillance to allow their growth and evolution. Nowhere is this as apparent as in multiple myeloma, a cancer of antibody-producing plasma cells, where a complex interplay between neoplastic cells and the immune microenvironment is required for the development and progression of disease. Decades of research has led to the discovery of a number of therapeutic agents, from cytotoxic drugs to genetically engineered cells that mediate their antimyeloma effects at least partially through altering these immune interactions. In this review, we discuss the history of immunotherapy and current practices in multiple myeloma, as well as the advances that promise to one day offer a cure for this deadly disease.

  View details for DOI 10.1158/1078-0432.CCR-20-3600

  View details for Web of Science ID 000753846600004

  View details for PubMedID 33771856

  View details for PubMedCentralID PMC8364865

• Clonal hematopoiesis in patients receiving chimeric antigen receptor T-cell therapy. Blood advances Miller, P. G., Sperling, A. S., Brea, E. J., Leick, M. B., Fell, G. G., Jan, M., Gohil, S. H., Tai, Y., Munshi, N. C., Wu, C. J., Neuberg, D. S., Maus, M. V., Jacobson, C., Gibson, C. J., Ebert, B. L. 2021; 5 (15): 2982-2986

  Abstract

  Chimeric antigen receptor (CAR) T-cells have emerged as an efficacious modality in patients with non-Hodgkin lymphoma (NHL) and multiple myeloma (MM). Clonal hematopoiesis of indeterminate potential (CHIP), a state in which mutations in hematopoietic cells give rise to a clonal population of cells, is more common in patients exposed to cytotoxic therapies, has been shown to influence inflammatory immune programs, and is associated with an adverse prognosis in patients with NHL and MM receiving autologous transplantation. We therefore hypothesized that CHIP could influence clinical outcomes in patients receiving CAR T-cell therapy. In a cohort of 154 patients with NHL or MM receiving CAR T-cells, we found that CHIP was present in 48% of patients and associated with increased rates of complete response and cytokine release syndrome severity, but only in patients younger than age 60 years. Despite these differences, CHIP was not associated with a difference in progression-free or overall survival, regardless of age. Our data suggest that CHIP can influence CAR T-cell biology and clinical outcomes, but, in contrast to autologous transplantation, CHIP was not associated with worse survival and should not be a reason to exclude individuals from receiving this potentially life-prolonging treatment.

  View details for DOI 10.1182/bloodadvances.2021004554

  View details for PubMedID 34342642

• Cancer therapies based on targeted protein degradation - lessons learned with lenalidomide NATURE REVIEWS CLINICAL ONCOLOGY Jan, M., Sperling, A. S., Ebert, B. L. 2021; 18 (7): 401-417

  Abstract

  For decades, anticancer targeted therapies have been designed to inhibit kinases or other enzyme classes and have profoundly benefited many patients. However, novel approaches are required to target transcription factors, scaffolding proteins and other proteins central to cancer biology that typically lack catalytic activity and have remained mostly recalcitrant to drug development. The selective degradation of target proteins is an attractive approach to expand the druggable proteome, and the selective oestrogen receptor degrader fulvestrant served as an early example of this concept. Following a long and tragic history in the clinic, the immunomodulatory imide drug (IMiD) thalidomide was discovered to exert its therapeutic activity via a novel and unexpected mechanism of action: targeting proteins to an E3 ubiquitin ligase for subsequent proteasomal degradation. This discovery has paralleled and directly catalysed myriad breakthroughs in drug development, leading to the rapid maturation of generalizable chemical platforms for the targeted degradation of previously undruggable proteins. Decades of clinical experience have established front-line roles for thalidomide analogues, including lenalidomide and pomalidomide, in the treatment of haematological malignancies. With a new generation of 'degrader' drugs currently in development, this experience provides crucial insights into class-wide features of degraders, including a unique pharmacology, mechanisms of resistance and emerging therapeutic opportunities. Herein, we review these past experiences and discuss their application in the clinical development of novel degrader therapies.

  View details for DOI 10.1038/s41571-021-00479-z

  View details for Web of Science ID 000624379400002

  View details for PubMedID 33654306

  View details for PubMedCentralID PMC8903027

• Biallelic loss of BCMA as a resistance mechanism to CAR T cell therapy in a patient with multiple myeloma NATURE COMMUNICATIONS Samur, M., Fulciniti, M., Samur, A., Bazarbachi, A., Tai, Y., Prabhala, R., Alonso, A., Sperling, A. S., Campbell, T., Petrocca, F., Hege, K., Kaiser, S., Loiseau, H., Anderson, K. C., Munshi, N. C. 2021; 12 (1): 868

  Abstract

  BCMA targeting chimeric antigen receptor (CAR) T cell therapy has shown deep and durable responses in multiple myeloma. However, relapse following therapy is frequently observed, and mechanisms of resistance remain ill-defined. Here, we perform single cell genomic characterization of longitudinal samples from a patient who relapsed after initial CAR T cell treatment with lack of response to retreatment. We report selection, following initial CAR T cell infusion, of a clone with biallelic loss of BCMA acquired by deletion of one allele and a mutation that creates an early stop codon on the second allele. This loss leads to lack of CAR T cell proliferation following the second infusion and is reflected by lack of soluble BCMA in patient serum. Our analysis suggests the need for careful detection of BCMA gene alterations in multiple myeloma cells from relapse following CAR T cell therapy.

  View details for DOI 10.1038/s41467-021-21177-5

  View details for Web of Science ID 000625204500023

  View details for PubMedID 33558511

  View details for PubMedCentralID PMC7870932

• Clonal hematopoiesis is associated with adverse outcomes in multiple myeloma patients undergoing transplant NATURE COMMUNICATIONS Mouhieddine, T. H., Sperling, A. S., Redd, R., Park, J., Leventhal, M., Gibson, C. J., Manier, S., Nassar, A. H., Capelletti, M., Huynh, D., Bustoros, M., Sklavenitis-Pistofidis, R., Tahri, S., Hornburg, K., Dumke, H., Itani, M. M., Boehner, C. J., Liu, C., AlDubayan, S. H., Reardon, B., Van Allen, E. M., Keats, J. J., Stewart, C., Mehr, S., Auclair, D., Schlossman, R. L., Munshi, N. C., Anderson, K. C., Steensma, D. P., Laubach, J. P., Richardson, P. G., Ritz, J., Ebert, B. L., Soiffer, R. J., Trippa, L., Getz, G., Neuberg, D. S., Ghobrial, I. M. 2020; 11 (1): 2996

  Abstract

  Multiple myeloma (MM) is a plasma-cell neoplasm that is treated with high-dose chemotherapy, autologous stem cell transplant (ASCT) and long-term immunomodulatory drug (IMiD) maintenance. The presence of somatic mutations in the peripheral blood is termed clonal hematopoiesis of indeterminate potential (CHIP) and is associated with adverse outcomes. Targeted sequencing of the stem cell product from 629 MM patients treated by ASCT at the Dana-Farber Cancer Institute (2003-2011) detects CHIP in 136/629 patients (21.6%). The most commonly mutated genes are DNMT3A, TET2, TP53, ASXL1 and PPM1D. Twenty-one from fifty-six patients (3.3%) receiving first-line IMiD maintenance develop a therapy-related myeloid neoplasm (TMN). However, regardless of CHIP status, the use of IMiD maintenance associates with improved PFS and OS. In those not receiving IMiD maintenance, CHIP is associated with decreased overall survival (OS) (HR:1.34, p = 0.02) and progression free survival (PFS) (HR:1.45, p < 0.001) due to an increase in MM progression.

  View details for DOI 10.1038/s41467-020-16805-5

  View details for Web of Science ID 000542758700026

  View details for PubMedID 32533060

  View details for PubMedCentralID PMC7293239

• Myelodysplastic syndromes (MDS) occurring in Agent Orange exposed individuals carry a mutational spectrum similar to that of <i>de novo</i> MDS LEUKEMIA & LYMPHOMA Sperling, A. S., Leventhal, M., Gibson, C. J., Ebert, B. L., Steensma, D. P. 2020; 61 (3): 728-731

  View details for DOI 10.1080/10428194.2019.1689394

  View details for Web of Science ID 000495941400001

  View details for PubMedID 31714164

  View details for PubMedCentralID PMC7268906

• Patterns of substrate affinity, competition, and degradation kinetics underlie biological activity of thalidomide analogs BLOOD Sperling, A. S., Burgess, M., Keshishian, H., Gasser, J. A., Bhatt, S., Jan, M., Slabicki, M., Sellar, R. S., Fink, E. C., Miller, P. G., Liddicoat, B. J., Sievers, Q. L., Sharma, R., Adams, D. N., Olesinski, E. A., Fulciniti, M., Udeshi, N. D., Kuhn, E., Letai, A., Munshi, N. C., Carr, S. A., Ebert, B. L. 2019; 134 (2): 160-170

  Abstract

  Pharmacologic agents that modulate ubiquitin ligase activity to induce protein degradation are a major new class of therapeutic agents, active in a number of hematologic malignancies. However, we currently have a limited understanding of the determinants of activity of these agents and how resistance develops. We developed and used a novel quantitative, targeted mass spectrometry (MS) assay to determine the relative activities, kinetics, and cell-type specificity of thalidomide and 4 analogs, all but 1 of which are in clinical use or clinical trials for hematologic malignancies. Thalidomide analogs bind the CRL4CRBN ubiquitin ligase and induce degradation of particular proteins, but each of the molecules studied has distinct patterns of substrate specificity that likely underlie the clinical activity and toxicities of each drug. Our results demonstrate that the activity of molecules that induce protein degradation depends on the strength of ligase-substrate interaction in the presence of drug, the levels of the ubiquitin ligase, and the expression level of competing substrates. These findings highlight a novel mechanism of resistance to this class of drugs mediated by competition between substrates for access to a limiting pool of the ubiquitin ligase. We demonstrate that increased expression of a nonessential substrate can lead to decreased degradation of other substrates that are critical for antineoplastic activity of the drug, resulting in drug resistance. These studies provide general rules that govern drug-dependent substrate degradation and key differences between thalidomide analog activity in vitro and in vivo.

  View details for DOI 10.1182/blood.2019000789

  View details for Web of Science ID 000474864100012

  View details for PubMedID 31043423

  View details for PubMedCentralID PMC6624968

• The genetics of myelodysplastic syndrome: from clonal haematopoiesis to secondary leukaemia NATURE REVIEWS CANCER Sperling, A. S., Gibson, C. J., Ebert, B. L. 2017; 17 (1): 5-19

  Abstract

  Myelodysplastic syndrome (MDS) is a clonal disease that arises from the expansion of mutated haematopoietic stem cells. In a spectrum of myeloid disorders ranging from clonal haematopoiesis of indeterminate potential (CHIP) to secondary acute myeloid leukaemia (sAML), MDS is distinguished by the presence of peripheral blood cytopenias, dysplastic haematopoietic differentiation and the absence of features that define acute leukaemia. More than 50 recurrently mutated genes are involved in the pathogenesis of MDS, including genes that encode proteins involved in pre-mRNA splicing, epigenetic regulation and transcription. In this Review we discuss the molecular processes that lead to CHIP and further clonal evolution to MDS and sAML. We also highlight the ways in which these insights are shaping the clinical management of MDS, including classification schemata, prognostic scoring systems and therapeutic approaches.

  View details for DOI 10.1038/nrc.2016.112

  View details for Web of Science ID 000391518900004

  View details for PubMedID 27834397

  View details for PubMedCentralID PMC5470392

• Differential and limited expression of mutant alleles in multiple myeloma BLOOD Rashid, N. U., Sperling, A. S., Bolli, N., Wedge, D. C., Van Loo, P., Tai, Y., Shammas, M. A., Fulciniti, M., Samur, M. K., Richardson, P. G., Magrangeas, F., Minvielle, S., Futreal, P., Anderson, K. C., Avet-Loiseau, H., Campbell, P. J., Parmigiani, G., Munshi, N. C. 2014; 124 (20): 3110-3117

  Abstract

  Recent work has delineated mutational profiles in multiple myeloma and reported a median of 52 mutations per patient, as well as a set of commonly mutated genes across multiple patients. In this study, we have used deep sequencing of RNA from a subset of these patients to evaluate the proportion of expressed mutations. We find that the majority of previously identified mutations occur within genes with very low or no detectable expression. On average, 27% (range, 11% to 47%) of mutated alleles are found to be expressed, and among mutated genes that are expressed, there often is allele-specific expression where either the mutant or wild-type allele is suppressed. Even in the absence of an overall change in gene expression, the presence of differential allelic expression within malignant cells highlights the important contribution of RNA-sequencing in identifying clinically significant mutational changes relevant to our understanding of myeloma biology and also for therapeutic applications.

  View details for DOI 10.1182/blood-2014-04-569327

  View details for Web of Science ID 000347462700015

  View details for PubMedID 25237203

  View details for PubMedCentralID PMC4231420

• Topoisomerase II binds nucleosome-free DNA and acts redundantly with topoisomerase I to enhance recruitment of RNA Pol II in budding yeast PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Sperling, A. S., Jeong, K., Kitada, T., Grunstein, M. 2011; 108 (31): 12693-12698

  Abstract

  DNA topoisomerases are believed to promote transcription by removing excessive DNA supercoils produced during elongation. However, it is unclear how topoisomerases in eukaryotes are recruited and function in the transcription pathway in the context of nucleosomes. To address this problem we present high-resolution genome-wide maps of one of the major eukaryotic topoisomerases, Topoisomerase II (Top2) and nucleosomes in the budding yeast, Saccharomyces cerevisiae. Our data indicate that at promoters Top2 binds primarily to DNA that is nucleosome-free. However, although nucleosome loss enables Top2 occupancy, the opposite is not the case and the loss of Top2 has little effect on nucleosome density. We also find that Top2 is involved in transcription. Not only is Top2 enriched at highly transcribed genes, but Top2 is required redundantly with Top1 for optimal recruitment of RNA polymerase II at their promoters. These findings and the examination of candidate-activated genes suggest that nucleosome loss induced by nucleosome remodeling factors during gene activation enables Top2 binding, which in turn acts redundantly with Top1 to enhance recruitment of RNA polymerase II.

  View details for DOI 10.1073/pnas.1106834108

  View details for Web of Science ID 000293385700036

  View details for PubMedID 21771901

  View details for PubMedCentralID PMC3150916

• Chromatin-dependent binding of the <i>S</i>. <i>cerevisiae</i> HMGB protein Nhp6A affects nucleosome dynamics and transcription GENES & DEVELOPMENT Dowell, N. L., Sperling, A. S., Mason, M. J., Johnson, R. C. 2010; 24 (18): 2031-2042

  Abstract

  The Saccharomyces cerevisiae protein Nhp6A is a model for the abundant and multifunctional high-mobility group B (HMGB) family of chromatin-associated proteins. Nhp6A binds DNA in vitro without sequence specificity and bends DNA sharply, but its role in chromosome biology is poorly understood. We show by whole-genome chromatin immunoprecipitation (ChIP) and high-resolution whole-genome tiling arrays (ChIP-chip) that Nhp6A is localized to specific regions of chromosomes that include ∼23% of RNA polymerase II promoters. Nhp6A binding functions to stabilize nucleosomes, particularly at the transcription start site of these genes. Both genomic binding and transcript expression studies point to functionally related groups of genes that are bound specifically by Nhp6A and whose transcription is altered by the absence of Nhp6. Genomic analyses of Nhp6A mutants specifically defective in DNA bending reveal a critical role of DNA bending for stabilizing chromatin and coregulation of transcription but not for targeted binding by Nhp6A. We conclude that the chromatin environment, not DNA sequence recognition, localizes Nhp6A binding, and that Nhp6A stabilizes chromatin structure and coregulates transcription.

  View details for DOI 10.1101/gad.1948910

  View details for Web of Science ID 000281815500006

  View details for PubMedID 20844014

  View details for PubMedCentralID PMC2939365

• Histone H3 N-terminus regulates higher order structure of yeast heterochromatin PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Sperling, A. S., Grunstein, M. 2009; 106 (32): 13153-13159

  Abstract

  In budding yeast, telomeres and the mating type (HM) loci are found in a heterochromatin-like silent structure initiated by Rap1 and extended by the interaction of Silencing Information Regulator (Sir) proteins with histones. Binding data demonstrate that both the H3 and H4 N-terminal domains required for silencing in vivo interact directly with Sir3 and Sir4 in vitro. The role of H4 lysine 16 deacetylation is well established in Sir3 protein recruitment; however, that of the H3 N-terminal tail has remained unclear. To characterize the role of H3 in silent chromatin formation and compare it to H4 we have generated comprehensive high resolution genome-wide binding maps of heterochromatin proteins. We found that H4 lysine 16 deacetylation is required for the recruitment and spreading of heterochromatin proteins at all telomeres and HM loci. In contrast, the H3 N terminus is required for neither recruitment nor spreading of Sir proteins. Instead, deletion of the H3 tail leads to increased accessibility within heterochromatin of an ectopic bacterial dam methylase and the decreased mobility of an HML heterochromatic fragment in sucrose gradients. These findings indicate an altered chromatin structure. We propose that Sir proteins recruited by the H4 tail then interact with the H3 tail to form a higher order silent chromatin structure.

  View details for DOI 10.1073/pnas.0906866106

  View details for Web of Science ID 000268877300004

  View details for PubMedID 19666585

  View details for PubMedCentralID PMC2726386

• Genetic screens for enhancers of brahma reveal functional interactions between the BRM chromatin-remodeling complex and the Delta-Notch signal transduction pathway in Drosophila GENETICS Armstrong, J. A., Sperling, A. S., Deuring, R., Manning, L., Moseley, S. L., Papoulas, O., Piatek, C. I., Doe, C. Q., Tamkun, J. W. 2005; 170 (4): 1761-1774

  Abstract

  The Drosophila trithorax group gene brahma (brm) encodes the ATPase subunit of a 2-MDa chromatin-remodeling complex. brm was identified in a screen for transcriptional activators of homeotic genes and subsequently shown to play a global role in transcription by RNA polymerase II. To gain insight into the targeting, function, and regulation of the BRM complex, we screened for mutations that genetically interact with a dominant-negative allele of brm (brm(K804R)). We first screened for dominant mutations that are lethal in combination with a brm(K804R) transgene under control of the brm promoter. In a distinct but related screen, we identified dominant mutations that modify eye defects resulting from expression of brm(K804R) in the eye-antennal imaginal disc. Mutations in three classes of genes were identified in our screens: genes encoding subunits of the BRM complex (brm, moira, and osa), other proteins directly involved in transcription (zerknullt and RpII140), and signaling molecules (Delta and vein). Expression of brm(K804R) in the adult sense organ precursor lineage causes phenotypes similar to those resulting from impaired Delta-Notch signaling. Our results suggest that signaling pathways may regulate the transcription of target genes by regulating the activity of the BRM complex.

  View details for DOI 10.1534/genetics.105.041327

  View details for Web of Science ID 000232033300027

  View details for PubMedID 15944353

  View details for PubMedCentralID PMC1449748

• Ciltacabtagene autoleucel in high-risk smoldering multiple myeloma: the CAR-PRISM phase 2 trial NATURE MEDICINE Nadeem, O., dos Santos, D., Nikiforow, S., Bosch-Vilaseca, A., O'Donnell, E., Redd, R., DeBraganca, K. C., Sperling, A. S., Liu, Y., McEntire, C., Arters, F., O'Donnell, C., Kineavy, B., Marto, M., Bergeron, A., Swenson, E., McHugh, K., Caron, A., Berry, Q., Wei, H., Durlacher, E., Grimm, E., Corrado, F., Bidikian, N., de Oca, R., Lengil, T., De Wiest, D., Gervais, C., Panaro, K., Smith, E. L., Anderson, K., Jacobson, C., Munshi, N. C., Richardson, P., Madduri, D., Schecter, J. M., Tendler, C., Wildgust, M. A., Trippa, L., Mateos, M., Ritz, J., Ghobrial, I. M. 2026

  Abstract

  High-risk smoldering multiple myeloma (HR-SMM) carries an increased risk of progression to multiple myeloma, making it an ideal setting to test whether chimeric antigen receptor (CAR) T cell therapy can achieve curative outcomes. Here in this phase 2 study, patients with HR-SMM received ciltacabtagene autoleucel (cilta-cel) at 0.3-0.5 × 106 or >0.5 × 106 viable CAR+ T cells per kilogram without induction or bridging therapy. Patients with >40% marrow involvement were excluded. Primary endpoints were dose-limiting toxicities (DLTs) and treatment-emergent adverse events; secondary endpoints included response and minimal residual disease (MRD) negativity. As of 11 February 2026, 20 patients had been treated. The trial met the prespecified endpoints. No DLTs occurred. Adverse events included transient cytopenias (90% grade 3/4) and cytokine release syndrome (100% grade 1/2). Non-immune effector cell-associated neurotoxicity syndrome neurologic toxicities (NINTs) occurred in seven patients, with four comprising cranial nerve palsies that completely resolved. Three patients had persistent grade 1 symptoms. At a median follow-up of 15.3 months, all patients achieved MRD negativity 10-6 by 2 months and have remained MRD negative. Sixteen patients with follow-up >6 months achieved a complete response; no progression or deaths were observed. Cilta-cel produced rapid, deep, sustained MRD-negative responses in HR-SMM without induction therapy. Toxicities were consistent with the safety profile of cilta-cel. ClinicalTrials.gov: NCT05767359 .

  View details for DOI 10.1038/s41591-026-04365-y

  View details for Web of Science ID 001744217900001

  View details for PubMedID 42010117

  View details for PubMedCentralID 7567803

• Genomically Smoldering Multiple Myeloma Is Not a Distinct Entity But a Collection of Monoclonal Gammopathy of Undetermined Significance or Multiple Myeloma JOURNAL OF CLINICAL ONCOLOGY Aktas Samur, A., Corre, J., Talluri, S., Shah, P., Graffeuil, A., Rivera, J., Fan, Y., Dakiki Korucu, B., Szalat, R., Fulciniti, M., Anderson, K. C., Sperling, A., Parmigiani, G., Avet-Loiseau, H., Munshi, N. C., Samur, M. 2026; 44 (4): 321-334

  Abstract

  The diagnosis of smoldering multiple myeloma (SMM) primarily relies on clinical features such as plasma cell involvement, immunoglobulin protein levels, and end-organ damage. However, as early intervention becomes a priority, the role of genomic features in differentiating risk is gaining attention.This study analyzed next-generation sequencing data from 224 precursor condition samples with 51 patients having paired SMM and multiple myeloma (MM) and 1,779 samples from newly diagnosed MM to identify genomic features linked to progression in SMM and those with a low-risk nonprogressor precursor condition.Our findings from paired samples revealed no significant differences in somatic alterations and clonal structures between SMM and MM samples from the same patient. This indicates that plasma cells in progressor SMM are genomically pre-equipped with changes that define myeloma. Over 80% of driver mutations were present at both time points, and more than 66% of progressor samples showed only minor clonal changes. We further compared genomic changes between nonprogressor and progressor SMM. Nonprogressor plasma cells showed significantly lower mutational load and the absence of copy number alterations on chromosome 8. They reduced focal genomic loss compared with progressor plasma cells. A scoring system using genomic features predictively identified patients with low-risk SMM unlikely to progress, validated on 101 additional independent samples, and additive clinical value of genomic classification was further shown in combination with 20/2/20.In summary, the genomic distinctions now suggest that a proportion of SMMs with progressor phenotype are akin to MM, whereas nonprogressor SMM has monoclonal gammopathy of undetermined significance-like characteristics. The results should influence further investigation in larger studies to inform future diagnostic criteria and trial designs.

  View details for DOI 10.1200/JCO-25-00289

  View details for Web of Science ID 001669157700004

  View details for PubMedID 41380103

• ID2 Suppresses Multiple Myeloma Cell Proliferation by Repressing the Activity of the Transcription Factor TCF3 BLOOD CANCER DISCOVERY Fulciniti, M., Yao, Y., Perini, T., Fong Ng, J., Schavgoulidze, A., Deng, S., Cui, J., Encinas Mayoral, J., Ladisa, F., Young, R. M., Epstein, C. B., White, C. M., Ott, C. J., Gulla, A., Matulis, S. M., Sperling, A. S., Morelli, E., Boise, L. H., Binder, M., Szalat, R., Samur, M. K., Anderson, K. C., Munshi, N. C. 2026; 7 (1): 129-141

  Abstract

  Transcription factors and their cofactors are major and selective nononcogene dependencies in multiple myeloma cells. By performing a gain-of-function perturbation screen in human multiple myeloma cell lines, we identified the inhibitor of DNA binding (ID) genes as putative suppressors of multiple myeloma cell fitness. Among them, ID2 was found to be downregulated in multiple myeloma patient cells and acted as a tumor suppressor by directly binding and repressing the basic helix-loop-helix factor TCF3, also known as E2A. Lower ID2 expression in multiple myeloma cells conferred a proliferative advantage by increasing TCF3 activity, leading to a dependency on this transcription factor. In contrast, ID2 overexpression reduced TCF3 binding to DNA, which resulted in cell-cycle arrest and a halt in multiple myeloma cell proliferation. The myeloma bone marrow milieu supported this process by further decreasing the expression of ID2 and enhancing TCF3 activity, partly via IL6, revealing a mechanism by which the tumor microenvironment affects multiple myeloma cell behavior.Multiple myeloma cells exploit the oncogenic and proliferative potential of TCF3 by downregulating the transcriptional regulator ID2, a process facilitated by the bone marrow microenvironment.

  View details for DOI 10.1158/2643-3230.BCD-25-0048

  View details for Web of Science ID 001660232000006

  View details for PubMedID 41104834

  View details for PubMedCentralID PMC12616536

• Putative multiple myeloma susceptibility genes identified by exome sequencing of 347 familial and early-onset cases LEUKEMIA Pertesi, M., Demangel, D., Niroula, A., Perrial, E., Escobar, M., Vallee, M., Liacos, C., Samur, M. K., Sperling, A. S., Munshi, N. C., Kastritis, E., Dimopoulos, M. A., McKay, J. D., Mellqvist, U., Hansson, M., Dumontet, C., Intergrp Francophone Myelome IFM, Nilsson, B. 2026; 40 (1): 235-240

  Abstract

  Multiple myeloma (MM) is the second most common blood malignancy, with several lines of evidence supporting an inherited genetic component. Here, we sequenced 177 affected individuals from 128 families, and 170 early-onset MM cases diagnosed before 55 years of age. Samples were identified and collected through nationwide efforts in France, Sweden, and Greece. We focused on rare germline protein truncating and likely deleterious missense variants in genes harboring variants in at least two families showing variant-disease segregation, and in additional index (≥2) and/or early-onset (≥2) cases. We identified likely pathogenic variants in ATM (N = 12), ANGPTL6 (N = 5), and FBXW9 (N = 6). Additionally, we detected variants in previously reported MM predisposition genes, including DIS3, EP300, and KDM1A. Our results represent the largest sequencing study on familial and early-onset MM to date, and further illuminate the constitutional genetic basis of MM.

  View details for DOI 10.1038/s41375-025-02802-4

  View details for Web of Science ID 001618322500001

  View details for PubMedID 41254116

  View details for PubMedCentralID 11300596

• Mezigdomide for multiple myeloma: a focus on phase 2 trial data EXPERT OPINION ON EMERGING DRUGS Mo, C. C., Liu, Y., Nadeem, O., Midha, S., Laubach, J. P., Salman, T., Nicholson, T., Donnelly, K., Bianchi, G., Sperling, A. S., Hartley-Brown, M. A., Richardson, P. G. 2025; 30 (3): 209-219

  Abstract

  The treatment armamentarium for multiple myeloma (MM) has evolved substantially over the past 20 years. Standard-of-care regimens for newly diagnosed MM and early relapsed/refractory disease (RRMM) include quadruplets and triplets comprising CD38 monoclonal antibodies, proteasome inhibitors, and/or immunomodulatory drugs, plus dexamethasone. Targeted agents and immune-based therapies are being used increasingly early in the treatment algorithm. There is an ongoing need for novel treatment options to improve outcomes with existing therapies and in subsequent lines of treatment.We review preclinical/clinical data on the cereblon E3 ligase modulator mezigdomide, which is in phase 3 investigation in RRMM. We searched the published literature using PubMed, plus congress abstracts from the past 5 years and current records on ClinicalTrials.gov, using the terms 'mezigdomide' or 'CC-92480' and 'myeloma.'Mezigdomide, which is not currently approved for the treatment of MM, has higher cereblon binding affinity and greater potency for substrate protein degradation than the immunomodulatory drugs, and this is translating into notable clinical efficacy in early-phase trials, including in poor-prognosis settings such as triple-class-refractory disease. It has shown synergistic effects in preclinical studies with standard-of-care therapies and is being evaluated clinically in various combinations, including with/following T-cell engaging therapies for RRMM.

  View details for DOI 10.1080/14728214.2025.2575570

  View details for Web of Science ID 001605315600001

  View details for PubMedID 41163399

• Expanding the druggable zinc-finger proteome defines properties of drug-induced degradation. Molecular cell Słabicki, M., Park, J., Nowak, R. P., Roy Burman, S. S., Pellman, J., Zou, C., Razumkov, H., Carreiro, J., Rastogi, S., Goldstein, A., Nagiec, M. M., Donovan, K. A., Che, J., Hunkeler, M., Geng, Q., Hsu, C. L., Lakshminarayan, M., Shu, C., Zon, R. L., Kozicka, Z., Park, P. M., Tsai, J. M., Yoon, H., Jones, L. H., Sperling, A. S., Gray, N. S., Fischer, E. S., Ebert, B. L. 2025; 85 (16): 3184-3201.e14

  Abstract

  Glutarimide analogs, such as thalidomide, redirect the E3 ubiquitin ligase CRL4CRBN to induce degradation of certain zinc finger (ZF) proteins. Although the core structural motif recognized by CRBN has been characterized, it does not fully explain substrate specificity. To explore the role of residues adjacent to this core motif, we constructed a comprehensive ZF reporter library of 9,097 reporters derived from 1,655 human ZF proteins and conducted a library-on-library screen with 29 glutarimide analogs to identify compounds that collectively degrade 38 ZF reporters. Cryo-electron microscopy and crystal structures of ZFs in complex with CRBN revealed the importance of interactions beyond the core ZF degron. We used systematic mutagenesis of ZFs and CRBN to identify modes of neosubstrate recruitment requiring distinct amino acids. Finally, we found subtle chemical variations in glutarimide analogs that alter target scope and selectivity, thus providing a roadmap for their rational design.

  View details for DOI 10.1016/j.molcel.2025.07.019

  View details for PubMedID 40845806

• International Myeloma Society/International Myeloma Working Group Consensus Recommendations on the Definition of High-Risk Multiple Myeloma JOURNAL OF CLINICAL ONCOLOGY Avet-Loiseau, H., Davies, F. E., Samur, M. K., Corre, J., D'Agostino, M., Kaiser, M. F., Raab, M. S., Weinhold, N., Gutierrez, N. C., Paiva, B., Neri, P., Weisel, K., Maura, F., Walker, B. A., Bustoros, M., Stewart, A., Usmani, S. Z., Hillengass, J., Chng, W., Keats, J. J., Martinez-Lopez, J., Sperling, A. S., Touzeau, C., Zhan, F., Raje, N. S., Cavo, M., Bolli, N., Ghobrial, I. M., Dhodapkar, M. V., Jagannath, S., Spencer, A., Parekh, S., Bahlis, N. J., Lonial, S., Sonneveld, P., Bergsagel, L., Orlowski, R. Z., Morgan, G., Mateos, M., Rajkumar, S., San Miguel, J. F., Anderson, K. C., Moreau, P., Kumar, S., Prosper, F., Munshi, N. C. 2025; 43 (24): 2739-2751

  Abstract

  Despite significant improvements in survival of patients with multiple myeloma (MM), outcomes remain heterogeneous, and a significant proportion of patients experience suboptimal outcomes. Importantly, traditional prognostic factors based on data from patients treated with older therapies no longer capture prognosis accurately in the contemporary era of novel triplet or quadruplet therapies. Therefore, risk stratification requires refinement in the context of available and investigational treatment options in routine practice and clinical trials, respectively. The current identification of high-risk MM (HRMM) in routine practice is based on the Revised International Staging System, which stratifies patients using a combination of widely available serum biomarkers and chromosomal abnormalities assessed via fluorescence in situ hybridization. In recent years, a substantial body of evidence concerning additional clinical, biological, and molecular/genomic prognostic factors has accumulated, along with new MM risk stratification tools and consensus reports. The International Myeloma Society, along with the International Myeloma Working Group, convened an Expert Panel with the primary aim of revisiting the definition of HRMM and formulating a practical and data-driven consensus definition, based on new evidence from molecular/genomic assays, updated clinical data, and contemporary risk stratification concepts. The Panel proposes the following Consensus Genomic Staging (CGS) of HRMM which relies upon the presence of at least one of these abnormalities: (1) del(17p), with a cutoff of >20% clonal fraction, and/or TP53 mutation; (2) an IgH translocation including t(4;14), t(14;16), or t(14;20) along with 1q+ and/or del(1p32); (3) monoallelic del(1p32) along with 1q+ or biallelic del(1p32); or (4) β2 microglobulin ≥5.5 mg/L with normal creatinine (<1.2 mg/dL).

  View details for DOI 10.1200/JCO-24-01893

  View details for Web of Science ID 001551329800001

  View details for PubMedID 40489728

• Invasive fungal disease is rare in patients with relapsed/refractory multiple myeloma treated with BCMA CAR T-cell therapy BLOOD ADVANCES Little, J. S., Pena, A., Kim, E., Yee, A. J., Nadeem, O., Midha, S., Sperling, A. S., Munshi, N. C., Raje, N., Frigault, M. J., Cirstea, D. D., Hammond, S. P. 2025; 9 (16): 4190-4194

  View details for DOI 10.1182/bloodadvances.2025016748

  View details for Web of Science ID 001604627800001

  View details for PubMedID 40561525

  View details for PubMedCentralID PMC12361759

• Bridging intensity is associated with impaired hematopoietic recovery after BCMA CAR-T therapy for multiple myeloma BLOOD ADVANCES Frenking, J. H., Zhou, X., Rejeski, K., Wagner, V., Costello, P., Hielscher, T., Gatti, L., Kauer, J., Nadeem, O., Mai, E. K., Michel, C. S., Friedrich, M. J., Sedloev, D., Weinhold, N., Goldschmidt, H., Herfarth, K., Schmitt, A., Hundemer, M., Schmitt, M., Muller-Tidow, C., Topp, M., Einsele, H., Dreger, P., Munshi, N. C., Sperling, A. S., Rasche, L., Sauer, S., Raab, M. S. 2025; 9 (16): 4151-4166

  Abstract

  Chimeric antigen receptor (CAR) T-cell (CAR-T) therapy represents a major advance in the treatment of relapsed/refractory multiple myeloma (RRMM). However, the long time span from leukapheresis to actual CAR-T infusion often necessitates bridging therapies. Because of limited knowledge about the effects of bridging on post-CAR-T clinical course and outcomes, the selection of treatment options is challenging. In this multicenter international observational study, we explored the impact of bridging therapy on hematopoietic reconstitution in 158 patients with RRMM treated with B-cell maturation antigen (BCMA)-directed CAR-T therapy. Based on exposure to classical cytotoxic (CTX) chemotherapy, we classified bridging regimens as non-CTX, intermediate CTX (1-2 CTX agents), or intensive CTX (≥3 CTX agents or high-dose therapy with stem cell transplantation). We found associations between the number of CTX agents used and impaired post-CAR-T hematopoietic reconstitution, evident across hematopoietic cell lineages and particularly manifesting during the late post-CAR-T period. Intensive CTX bridging was associated with a prolonged time to neutrophil and platelet recovery, distinct patterns of hematopoietic recovery (eg, an intermittent phenotype characterized by a second drop), an increased susceptibility to severe infections and a significantly increased risk for severe late cytopenias in univariate and multivariate models. Taken together, these results highlight that bridging intensity distinctly shapes the trajectory of hematopoietic recovery after BCMA CAR-T therapy. Targeted and novel immunotherapies could provide alternatives for bridging, and high-risk patients may particularly benefit from enhanced monitoring, prophylaxis, and supportive care.

  View details for DOI 10.1182/bloodadvances.2024015732

  View details for Web of Science ID 001603518400001

  View details for PubMedID 40267180

  View details for PubMedCentralID PMC12361772

• Two clones, one niche: how CH shapes the MM microenvironment BLOOD Sperling, A. S. 2025; 146 (5): 524-525

  View details for DOI 10.1182/blood.2025029174

  View details for Web of Science ID 001545631100004

  View details for PubMedID 40742726

  View details for PubMedCentralID PMC12333223

• Real-World Experience with Teclistamab for Relapsed/ Refractory Multiple Myeloma from the U.S. Myeloma Immunotherapy Consortium. Blood cancer discovery Razzo, B. M., Midha, S., Portuguese, A. J., Grajales-Cruz, A. F., De Menezes Silva Corraes, A., Costello, P., Liu, Y., Sperling, A. S., Nadeem, O., Dima, D., Banerjee, R., Cowan, A. J., Afrough, A., Anderson, L. D., Lieberman-Cribbin, A., Kaur, G., Goyal, A., Atrash, S., Ferreri, C. J., Voorhees, P. M., Pasvolsky, O., Lee, H. C., Patel, K. K., Julian, K. L., Forsberg, P. A., Herr, M. M., Chhabra, S., Parrondo, R. D., Lin, Y., Chen, A., Susanibar-Adaniya, S. P., Khouri, J., Raza, S., Anwer, F., Vazquez-Martinez, M., Castaneda Puglianini, O., Sborov, D. W., Davis, J. A., Rossi, A., Shune, L., Bhurtel, J., Hwang, W. T., Hansen, D. K., Sidana, S., Garfall, A. L., Richard, S. 2025

  Abstract

  Teclistamab is an anti-CD3xBCMA bispecific antibody approved for use in relapsed/refractory multiple myeloma (MM). We undertook a retrospective study of post-approval, real-world outcomes with teclistamab in the U.S. MM Immunotherapy Consortium. Among 509 patients, 89% would have been ineligible for the MajesTEC-1 trial, primarily due to prior BCMA-directed therapy, cytopenias, or diminished performance status. Cytokine release syndrome occurred in 54% (1.4% grade ≥3) and immune effector cell-associated neurotoxicity syndrome in 11% (2.2% grade ≥3) with no fatal events. Infections occurred in 42% and contributed to death in 5%. Partial response (PR) or better was achieved in 53% and very good PR (VGPR) or better in 45%. With 10.1 months median follow-up, estimated median progression-free survival (PFS) was 5.8 months, and 12-month overall survival was 61%. Independent predictors of

  View details for DOI 10.1158/2643-3230.BCD-24-0354

  View details for PubMedID 40629516

• Carfilzomib prescribing patterns and outcomes for relapsed or refractory multiple myeloma: a real-world analysis BLOOD CANCER JOURNAL Dong, S., Banerjee, R., Khan, A. M., Wang, M., Wang, X., Afghahi, A., Afrough, A., Janakiram, M., Wang, B., Cowan, A. J., Sperling, A. S., Anderson Jr, L. D., Rajkumar, S., Kaur, G. 2025; 15 (1): 48

  Abstract

  Despite the widespread use of carfilzomib (K) in relapsed/refractory multiple myeloma (RRMM), there is no consensus on optimal K dose in milligrams per square meter (mg/m2) or dosing schedule. We assessed three modern K prescribing patterns in RRMM using a large United States electronic health record-derived database. Our final cohort (n = 486) included 136 patients (28.0%) who received K 56 mg/m2 once weekly (K56-1x), 86 (17.7%) who received 56 mg/m2 twice weekly (K56-2x), and 264 (54.3%) who received 70 mg/m2 once weekly (K70-1x). Between 2016 and 2023, once-weekly dosing became more common: K70-1x proportions changed from 21.1% in 2016 to 50.6% in 2023, K56-1x from 15.8% to 37.0%, and K56-2x from 63.2% to 12.3%. Median progression-free survival was 13.0 months [95% confidence interval (CI) 11.2-20.7] for K56-1x, 13.2 months (95% CI 9.0-28.1 months) for K56-2x, and 10.9 months (95% CI 9.9-15.3 months) for K70-1x; these differences were not statistically significant (log-rank p = 0.46). Rates of heart failure was comparable (<5% in all cohorts). In summary, our findings do not support improved outcomes with twice-weekly carfilzomib in RRMM. K56-1x may provide the best balance of efficacy, safety, and avoidance of time toxicity from frequent infusions.

  View details for DOI 10.1038/s41408-025-01256-2

  View details for Web of Science ID 001455923800002

  View details for PubMedID 40155649

  View details for PubMedCentralID PMC11953251

• Cytopenias in BCMA CAR T: unraveling inflammatory mechanisms BLOOD ADVANCES Sperling, A. S. 2024; 8 (21): 5527-5528

  View details for DOI 10.1182/bloodadvances.2024014308

  View details for Web of Science ID 001346513100001

  View details for PubMedID 39446348

  View details for PubMedCentralID PMC11544301

• Targeting Ikaros and Aiolos: reviewing novel protein degraders for the treatment of multiple myeloma, with a focus on iberdomide and mezigdomide. Expert review of hematology Liu, Y., Mo, C. C., Hartley-Brown, M. A., Sperling, A. S., Midha, S., Yee, A. J., Bianchi, G., Piper, C., Tattersall, A., Nadeem, O., Laubach, J. P., Richardson, P. G. 2024; 17 (8): 445-465

  Abstract

  INTRODUCTION: The treatment of multiple myeloma (MM) is evolving rapidly. Quadruplet regimens incorporating proteasome inhibitors, immunomodulatory drugs (IMiDs), and CD38 monoclonal antibodies have emerged as standard-of-care options for newly diagnosed MM, and numerous novel therapies have been approved for relapsed/refractory MM. However, there remains a need for novel options in multiple settings, including refractoriness to frontline standards of care.AREAS COVERED: Targeting degradation of IKZF1 and IKZF3 - Ikaros and Aiolos - through modulation of cereblon, an E3 ligase substrate recruiter/receptor, is a key mechanism of action of the IMiDs and the CELMoD agents. Two CELMoD agents, iberdomide and mezigdomide, have demonstrated substantial preclinical and clinical activity in MM and have entered phase 3 investigation. Using a literature search methodology comprising searches of PubMed (unlimited time-frame) and international hematology/oncology conference abstracts (2019-2023), this paper reviews the importance of Ikaros and Aiolos in MM, the mechanism of action of the IMiDs and CELMoD agents and their relative potency for targeting Ikaros and Aiolos, and preclinical and clinical data on iberdomide and mezigdomide.EXPERT OPINION: Emerging data suggest that iberdomide and mezigdomide have promising activity, including in IMiD-resistant settings and, pending phase 3 findings, may provide additional treatment options for patients with MM.

  View details for DOI 10.1080/17474086.2024.2382897

  View details for PubMedID 39054911

• Round Table Discussion on Optimal Clinical Trial Design in Precursor Multiple Myeloma BLOOD CANCER DISCOVERY Ghobrial, I. M., Gormley, N., Kumar, S. K., Mateos, M., Bergsagel, P., Chesi, M., Dhodapkar, M. V., Dispenzieri, A., Fonseca, R., Getz, G., Kastritis, E., Kristinsson, S. Y., Martinez-Climent, J., Manier, S., Marinac, C. R., Maura, F., Morgan, G. J., Davies, F. E., Nadeem, O., Nuvolone, M., Paiva, B., O'Donnell, E., Prosper, F., Shah, U. A., Sklavenitis-Pistofidis, R., Sperling, A. S., Vassiliou, G. S., Munshi, N. C., Castle, P. E., Anderson, K. C., San Miguel, J. F. 2024; 5 (3): 146-152

  Abstract

  While the current approach to precursor hematologic conditions is to "watch and wait," this may change with the development of therapies that are safe and extend survival or delay the onset of symptomatic disease. The goal of future therapies in precursor hematologic conditions is to improve survival and prevent or delay the development of symptomatic disease while maximizing safety. Clinical trial considerations in this field include identifying an appropriate at-risk population, safety assessments, dose selection, primary and secondary trial endpoints including surrogate endpoints, control arms, and quality-of-life metrics, all of which may enable more precise benefit-risk assessment.

  View details for DOI 10.1158/2643-3230.BCD-24-0022

  View details for Web of Science ID 001245344800001

  View details for PubMedID 38441243

  View details for PubMedCentralID PMC11061588

• Acute Lymphoblastic Leukemia with Myeloid Mutations Is a High-Risk Disease Associated with Clonal Hematopoiesis BLOOD CANCER DISCOVERY Saygin, C., Zhang, P., Stauber, J., Aldoss, I., Sperling, A. S., Weeks, L. D., Luskin, M. R., Knepper, T. C., Wanjari, P., Wang, P., Lager, A. M., Fitzpatrick, C., Segal, J. P., Gharghabi, M., Gurbuxani, S., Venkataraman, G., Cheng, J. X., Eisfelder, B. J., Bohorquez, O., Patel, A. A., Umesh Nagalakshmi, S., Jayaram, S., Odenike, O. M., Larson, R. A., Godley, L. A., Arber, D. A., Gibson, C. J., Munshi, N. C., Marcucci, G., Ebert, B. L., Greally, J. M., Steidl, U., Lapalombella, R., Shah, B. D., Stock, W. 2024; 5 (3): 164-179

  Abstract

  Myeloid neoplasms arise from preexisting clonal hematopoiesis (CH); however, the role of CH in the pathogenesis of acute lymphoblastic leukemia (ALL) is unknown. We found that 18% of adult ALL cases harbored TP53, and 16% had myeloid CH-associated gene mutations. ALL with myeloid mutations (MyM) had distinct genetic and clinical characteristics, associated with inferior survival. By using single-cell proteogenomic analysis, we demonstrated that myeloid mutations were present years before the diagnosis of ALL, and a subset of these clones expanded over time to manifest as dominant clones in ALL. Single-cell RNA sequencing revealed upregulation of genes associated with cell survival and resistance to apoptosis in B-ALL with MyM, which responds better to newer immunotherapeutic approaches. These findings define ALL with MyM as a high-risk disease that can arise from antecedent CH and offer new mechanistic insights to develop better therapeutic and preventative strategies.CH is a precursor lesion for lymphoblastic leukemogenesis. ALL with MyM has distinct genetic and clinical characteristics, associated with adverse survival outcomes after chemotherapy. CH can precede ALL years before diagnosis, and ALL with MyM is enriched with activated T cells that respond to immunotherapies such as blinatumomab. See related commentary by Iacobucci, p. 142.

  View details for DOI 10.1158/2643-3230.BCD-23-0106

  View details for Web of Science ID 001245344800003

  View details for PubMedID 38150184

  View details for PubMedCentralID PMC11061587

• Cellular Mass Response to Therapy Correlates With Clinical Response for a Range of Malignancies JCO PRECISION ONCOLOGY Stevens, M. M., Kimmerling, R. J., Olcum, S., Vacha, M., Labella, R., Minnah, A., Katsis, K., Fujii, J., Shaheen, Z., Sundaresan, S., Criscitiello, J., Niesvizky, R., Raje, N., Branagan, A., Krishnan, A., Jagannath, S., Parekh, S., Sperling, A. S., Rosenbaum, C. A., Munshi, N., Luskin, M. R., Tamrazi, A., Reid, C. A. 2024; 8: e2300349

  Abstract

  Cancer patients with advanced-stage disease have poor prognosis, typically having limited options for efficacious treatment, and genomics-based therapy guidance continues to benefit only a fraction of patients. Next-generation ex vivo approaches, such as cell mass-based response testing (MRT), offer an alternative precision medicine approach for a broader population of patients with cancer, but validation of clinical feasibility and potential impact remain necessary.We evaluated the clinical feasibility and accuracy of using live-cell MRT to predict patient drug sensitivity. Using a unified measurement workflow with a 48-hour result turnaround time, samples were subjected to MRT after treatment with a panel of drugs in vitro. After completion of therapeutic course, clinical response data were correlated with MRT-based predictions of outcome. Specimens were collected from 104 patients with solid (n = 69) and hematologic (n = 35) malignancies, using tissue formats including needle biopsies, malignant fluids, bone marrow aspirates, and blood samples. Of the 81 (78%) specimens qualified for MRT, 41 (51%) patients receiving physician-selected therapies had treatments matched to MRT.MRT demonstrated high concordance with clinical responses with an odds ratio (OR) of 14.80 (P = .0003 [95% CI, 2.83 to 102.9]). This performance held for both solid and hematologic malignances with ORs of 20.67 (P = .0128 [95% CI, 1.45 to 1,375.57]) and 8.20 (P = .045 [95% CI, 0.77 to 133.56]), respectively. Overall, these results had a predictive accuracy of 80% (P = .0026 [95% CI, 65 to 91]).MRT showed highly significant correlation with clinical response to therapy. Routine clinical use is technically feasible and broadly applicable to a wide range of samples and malignancy types, supporting the need for future validation studies.

  View details for DOI 10.1200/PO.23.00349

  View details for Web of Science ID 001282878400007

  View details for PubMedID 38237098

  View details for PubMedCentralID PMC10805426

• Mezigdomide is effective alone and in combination with Menin inhibition in pre-clinical models of KMT2A-r and NPM1c AML. Blood Bourgeois, W., Cutler, J. A., Aubrey, B. J., Wenge, D., Perner, F., Martucci, C., Henrich, J. A., Klega, K., Nowak, R. P., Donovan, K. A., Boileau, M., Wen, Y., Hatton, C., Apazidis, A. A., Olsen, S. N., Kirmani, N., Pikman, Y., Pollard, J. A., Perry, J. A., Sperling, A. S., Ebert, B. L., McGeehan, G. M., Crompton, B. D., Fischer, E. S., Armstrong, S. A. 2023

  Abstract

  Small molecules that target the MENIN-KMT2A protein-protein interaction (Menin inhibitors) have recently entered clinical trials in lysine methyltransferase 2A (KMT2A, MLL1) rearranged (KMT2A-r) and nucleophosmin mutant (NPM1c) acute myeloid leukemia (AML) and are demonstrating encouraging results. However, rationally chosen combination therapy is needed to improve responses and prevent resistance. We have previously identified IKZF1/IKAROS as a target in KMT2A-r AML and shown in preclinical models that IKAROS protein degradation with lenalidomide or iberdomide has modest single-agent activity yet can synergize with Menin inhibitors. Recently, the novel IKAROS degrader mezigdomide was developed with greatly enhanced IKAROS protein degradation. In this study we show that mezigdomide has increased preclinical activity in vitro as a single-agent in KMT2A-r and NPM1c AML cell lines, including sensitivity in cell lines resistant to lenalidomide and iberdomide. Further, we demonstrate that mezigdomide has the greatest capacity to synergize with and induce apoptosis in combination with Menin inhibitors, including in MEN1 mutant models. We show that the superior activity of mezigdomide compared to lenalidomide or iberdomide is due to its increased depth, rate, and duration of IKAROS protein degradation. Single-agent mezigdomide was efficacious in five patient derived xenograft (PDX) models of KMT2A-r and one NPM1c AML. The combination of mezigdomide with the Menin inhibitor VTP-50469 increased survival and prevented and overcame MEN1 mutations that mediate resistance in patients receiving Menin inhibitor monotherapy. These results support prioritization of mezigdomide for early phase clinical trials in KMT2A-r and NPM1c AML, either as a single-agent or in combination with Menin inhibitors.

  View details for DOI 10.1182/blood.2023021105

  View details for PubMedID 38096371

• A safety review of recently approved and emerging drugs for patients with relapsed or refractory multiple myeloma EXPERT OPINION ON DRUG SAFETY Midha, S., Hartley-Brown, M. A., Mo, C. C., Hossain, S., Nadeem, O., O'Donnell, E. K., Bianchi, G., Sperling, A. S., Laubach, J. P., Richardson, P. G. 2023; 22 (11): 1049-1071

  Abstract

  Multiple new drugs have been approved over the past 5 years for the treatment of relapsed/refractory multiple myeloma (RRMM), and these are being increasingly widely used. Clinicians need to familiarize themselves with common toxicities associated with these drugs and with novel toxicities requiring specific management and supportive care.We review common toxicities associated with agents approved for RRMM in the past 5 years, including the anti-CD38 monoclonal antibody isatuximab, the antibody-drug conjugate belantamab mafodotin, the bispecific antibody teclistamab, the chimeric antigen receptor (CAR) T cell products idecabtagene vicleucel and ciltacabtagene autoleucel, the selective inhibitor of nuclear export compound selinexor, and the drug-peptide conjugate melflufen, as well as toxicities associated with emerging agents for RRMM including additional bispecific antibodies, the BCL-2 inhibitor venetoclax, and the cereblon E3 ligase modulators iberdomide and mezigdomide. We searched the published literature using PubMed, plus congress abstracts, for the above list of drug names or classes and 'myeloma.'Optimal management of toxicities associated with these recently approved and emerging therapies will be critical in maximizing clinical benefit and aiding widespread adoption in routine clinical practice. We summarize current recommendations and guidelines and provide expert insights into supportive care requirements.

  View details for DOI 10.1080/14740338.2023.2274420

  View details for Web of Science ID 001100411600001

  View details for PubMedID 37906232

• Respiratory infections predominate after day 100 following B-cell maturation antigen-directed CAR T-cell therapy BLOOD ADVANCES Little, J. S., Tandon, M., Hong, J., Nadeem, O., Sperling, A. S., Raje, N., Munshi, N., Frigault, M., Barmettler, S., Hammond, S. P. 2023; 7 (18): 5485-5495

  Abstract

  Infections are an important complication after B-cell maturation antigen (BCMA)-directed chimeric antigen receptor (CAR) T-cell therapy and risks may differ between the early and late periods. We evaluated infections in 99 adults who received a first BCMA-directed CAR T-cell therapy (commercial and investigational autologous BCMA CAR T-cell products at the recommended phase 2 dose) for relapsed/refractory multiple myeloma between November 2016 and May 2022. Infections were recorded until day 365, if patients experienced symptoms with a microbiologic diagnosis, or for symptomatic site-specific infections treated with antimicrobials. One-year cumulative incidence functions were calculated based on time to first respiratory infection using dates of infection-free death and receipt of additional antineoplastic therapies as competing risks. Secondary analysis evaluated risk factors for late respiratory infections using univariate and multivariable Cox regression models. Thirty-seven patients (37%) experienced 64 infectious events over the first year after BCMA-directed CAR T-cell therapy, with 42 early infectious events (days, 0-100), and 22 late infectious events (days, 101-365). Respiratory infections were the most common site-specific infection and the relative proportion of respiratory infections increased in the late period (31% of early events vs 77% of late events). On multivariable analysis, hypogammaglobulinemia (hazard ratio [HR], 6.06; P = .044) and diagnosis of an early respiratory viral infection (HR, 2.95; P = .048) were independent risk factors for late respiratory infection. Respiratory infections predominate after BCMA CAR T-cell therapy, particularly after day 100. Hypogammaglobulinemia and diagnosis of an early respiratory infection are risk factors for late respiratory infections that may be used to guide targeted preventive strategies.

  View details for DOI 10.1182/bloodadvances.2023010524

  View details for Web of Science ID 001078758600001

  View details for PubMedID 37486599

  View details for PubMedCentralID PMC10514400

• Selinexor: Targeting a novel pathway in multiple myeloma EJHAEM Mo, C. C., Yee, A. J., Midha, S., Hartley-Brown, M. A., Nadeem, O., O'Donnell, E. K., Bianchi, G., Sperling, A. S., Laubach, J. P., Richardson, P. G. 2023; 4 (3): 792-810

  Abstract

  Selinexor is an orally bioavailable selective inhibitor of nuclear export compound that inhibits exportin-1 (XPO1), a novel therapeutic target that is overexpressed in multiple myeloma (MM) and is responsible for the transport of ∼220 nuclear proteins to the cytoplasm, including tumour suppressor proteins. Inhibition of this process has demonstrated substantial antimyeloma activity in preclinical studies, both alone and in combination with established MM therapeutics. Based on a clinical trial programme encompassing multiple combination regimens, selinexor-based therapy has been approved for the treatment of relapsed/refractory MM (RRMM), with selinexor-dexamethasone approved in the later-relapse setting for penta-refractory patients and selinexor-bortezomib-dexamethasone approved for patients who have received ≥1 prior therapy. Here, we provide a comprehensive review of the clinical data on selinexor-based regimens, including recent updates from the 2022 American Society of Hematology annual meeting, and summarise ongoing studies of this novel targeted agent in newly diagnosed MM and RRMM.

  View details for DOI 10.1002/jha2.709

  View details for Web of Science ID 001228421200034

  View details for PubMedID 37601856

  View details for PubMedCentralID PMC10435704

• Clonal Hematopoiesis in Young Women Treated for Breast Cancer CLINICAL CANCER RESEARCH Gibson, C. J., Fell, G., Sella, T., Sperling, A. S., Snow, C., Rosenberg, S. M., Kirkner, G., Patel, A., Dillon, D., Bick, A. G., Neuberg, D., Partridge, A. H., Miller, P. G. 2023; 29 (13): 2551-2558

  Abstract

  Young women treated for breast cancer with cytotoxic therapies are at risk for clonal hematopoiesis of indeterminate potential (CHIP), a condition in which blood cells carrying a somatic mutation associated with hematologic malignancy comprise at least 4% of the total blood system. CHIP has primarily been studied in older patient cohorts with limited clinical phenotyping.We performed targeted sequencing on longitudinal blood samples to characterize the clonal hematopoietic landscape of 878 women treated for breast cancer enrolled in the prospective Young Women's Breast Cancer Study.We identified somatic driver mutations in 252 study subjects (28.7%), but only 24 (2.7%) had clones large enough to meet criteria for CHIP. The most commonly mutated genes were DNMT3A and TET2, similar to mutations observed in noncancer cohorts. At 9-year median follow-up, we found no association between the presence of a somatic blood mutation (regardless of clone size) and adverse breast cancer (distant relapse-free survival) or non-breast cancer-related outcomes in this cohort. A subset of paired blood samples obtained over 4 years showed no evidence of mutant clonal expansion, regardless of genotype. Finally, we identified a subset of patients with likely germline mutations in genes known to contribute to inherited cancer risk, such as TP53 and ATM.Our data show that for young women with early-stage breast cancer, CHIP is uncommon after cytotoxic exposure, is unlikely to contribute to adverse outcomes over the decade-long follow-up and may not require additional monitoring if discovered incidentally.

  View details for DOI 10.1158/1078-0432.CCR-23-0050

  View details for Web of Science ID 001026671900001

  View details for PubMedID 37115512

  View details for PubMedCentralID PMC10330424

• High-dose melphalan treatment significantly increases mutational burden at relapse in multiple myeloma BLOOD Samur, M., Roncador, M., Samur, A., Fulciniti, M., Bazarbachi, A., Szalat, R., Shammas, M. A., Sperling, A. S., Richardson, P. G., Magrangeas, F., Minvielle, S., Perrot, A., Corre, J., Moreau, P., Thakurta, A., Parmigiani, G., Anderson, K. C., Avet-Loiseau, H., Munshi, N. C. 2023; 141 (14): 1724-1736

  Abstract

  High-dose melphalan (HDM) improves progression-free survival in multiple myeloma (MM), yet melphalan is a DNA-damaging alkylating agent; therefore, we assessed its mutational effect on surviving myeloma cells by analyzing paired MM samples collected at diagnosis and relapse in the IFM 2009 study. We performed deep whole-genome sequencing on samples from 68 patients, 43 of whom were treated with RVD (lenalidomide, bortezomib, and dexamethasone) and 25 with RVD + HDM. Although the number of mutations was similar at diagnosis in both groups (7137 vs 7230; P = .67), the HDM group had significantly more mutations at relapse (9242 vs 13 383, P = .005). No change in the frequency of copy number alterations or structural variants was observed. The newly acquired mutations were typically associated with DNA damage and double-stranded breaks and were predominantly on the transcribed strand. A machine learning model, using this unique pattern, predicted patients who would receive HDM with high sensitivity, specificity, and positive prediction value. Clonal evolution analysis showed that all patients treated with HDM had clonal selection, whereas a static progression was observed with RVD. A significantly higher percentage of mutations were subclonal in the HDM cohort. Intriguingly, patients treated with HDM who achieved complete remission (CR) had significantly more mutations at relapse yet had similar survival rates as those treated with RVD who achieved CR. This similarity could have been due to HDM relapse samples having significantly more neoantigens. Overall, our study identifies increased genomic changes associated with HDM and provides rationale to further understand clonal complexity.

  View details for DOI 10.1182/blood.2022017094

  View details for Web of Science ID 000981234300001

  View details for PubMedID 36603186

  View details for PubMedCentralID PMC10273091

• Clonal hematopoiesis of indeterminate potential and risk of death from COVID-19 BLOOD Miller, P. G., Fell, G. G., Foy, B. H., Scherer, A. K., Gibson, C. J., Sperling, A. S., Burugula, B. B., Nakao, T., Uddin, M. M., Warren, H., Bry, L., Pozdnyakova, O., Frigault, M. J., Bick, A. G., Neuberg, D., Higgins, J. M., Mansour, M. K., Natarajan, P., Kim, A. S., Kitzman, J. O., Ebert, B. L. 2022; 140 (18): 1993-1997

  Abstract

  Two Letters to Blood address the risks of COVID-19 in populations with precursors of hematological disease. In the first article, Miller and colleagues report on whether clonal hematopoiesis of intermediate potential (CHIP) is associated with adverse outcomes with COVID-19, finding no association between CHIP and 28-day mortality while providing data indirectly linking IL-6 signaling and patient outcomes. In the second article, Ho and colleagues investigate the outcomes of patients with monoclonal gammopathy of undetermined significance (MGUS) with COVID-19, reporting that one-fourth had a severe infection and that on multivariable analysis, adverse outcomes are more likely if immunoparesis is present.

  View details for DOI 10.1182/blood.2022018052

  View details for Web of Science ID 000920497500010

  View details for PubMedID 36096050

  View details for PubMedCentralID PMC9474399

• Quality of life, psychological distress, and prognostic perceptions in patients with multiple myeloma CANCER O'Donnell, E. K., Shapiro, Y. N., Yee, A. J., Nadeem, O., Hu, B. Y., Laubach, J. P., Branagan, A. R., Anderson, K. C., Mo, C. C., Munshi, N. C., Ghobrial, I. M., Sperling, A. S., Agyemang, E. A., Burke, J. N., Harrington, C. C., Richardson, P. G., Raje, N. S., El-Jawahri, A. 2022; 128 (10): 1996-2004

  Abstract

  Multiple myeloma (MM) is an incurable hematologic malignancy requiring long-term, continuous therapy. Despite its chronic and unrelenting course, studies examining quality of life (QOL), psychological distress, and perceptions of prognosis by line of therapy are lacking.The authors conducted a cross-sectional, multisite study of patients undergoing treatment for MM (excluding maintenance) between June 2020 and January 2021. The authors conducted purposeful sampling and recruited patients to 3 cohorts based on lines of therapy: 1) newly diagnosed receiving first-line therapy; 2) 2 to 3 lines; and 3) 4 or more lines. Patients completed validated questionnaires to assess their QOL, fatigue, psychological distress, and perceptions of prognosis.A total of 180 patients with MM were enrolled (newly diagnosed [n = 60], 2 to 3 lines [n = 60], and ≥4 lines of therapy [n = 60]). QOL, symptom burden, and fatigue scores did not differ by lines of therapy. There were no statistically significant differences in psychological distress by line of therapy. The rates of clinically significant depression, anxiety, and post-traumatic stress disorder symptoms were 23.9% (43 of 180), 23.9% (43 of 180), and 24.4% (44 of 180), respectively. Most patients (84.7%, 149 of 176) reported that their oncologist told them their cancer was incurable, but only 30.6% (53 of 173) acknowledged that they were terminally ill, and 42.0% (73 of 174) reported that they thought their cancer was incurable.Patients with MM undergoing treatment experience impaired QOL and elevated psychological distress across the disease continuum, regardless of line of therapy. A substantial proportion of patients with MM have significant misperceptions about their prognosis and the curability of their illness despite reporting being informed of the prognosis by their oncologist.This study discusses 180 patients with MM (newly diagnosed [n = 60], 2-3 lines [n = 60], and ≥4 lines of therapy [n = 60]). Quality of life, symptom burden, and fatigue scores do not differ by lines of therapy. There are also no statistically significant differences in psychological distress by line of therapy. The rates of clinically significant depression, anxiety, and post-traumatic stress disorder symptoms are 23.9%, 23.9%, and 24.4%, respectively. Most patients (84.7%) report that their oncologist told them their cancer was incurable, but only 30.6% acknowledge that they are terminally ill, and 42.0% report that they thought their cancer was incurable.

  View details for DOI 10.1002/cncr.34134

  View details for Web of Science ID 000755186100001

  View details for PubMedID 35167125

• The emerging importance and evolving understanding of clonal hematopoiesis in multiple myeloma SEMINARS IN ONCOLOGY DeStefano, C. B., Gibson, S. J., Sperling, A. S., Richardson, P. G., Ghobrial, I., Mo, C. C. 2022; 49 (1): 19-26

  Abstract

  Multiple myeloma (MM) is the second most common hematologic malignancy diagnosed in the United States. With a growing arsenal of novel therapies, patients are living longer and hence are at increased risk of secondary cancers such as myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). While MDS-associated cytogenetic abnormalities have been described in patients with a diagnosis of for decades, clonal hematopoiesis (CH) has been described only recently. CH has been shown to correlate with inferior survival in MM due to increased risk of disease progression in patients who are treated with high-dose melphalan without lenalidomide maintenance. When involving specific high-risk genes, multiple genes, or when present at high variant allelic frequencies, CH could also potentially elevate the risk of secondary MDS and/or AML, cardiovascular events, and venous thromboembolic events. Despite growing knowledge about CH in patients with MM, many questions remain unanswered. Further studies are needed to better understand the prognostic and therapeutic significance of CH in MM and its precursor conditions, as well as the effect of specific treatments on long-term outcome.

  View details for DOI 10.1053/j.seminoncol.2022.01.009

  View details for Web of Science ID 000806193300004

  View details for PubMedID 35105451

• Functional dissection of inherited non-coding variation influencing multiple myeloma risk. Nature communications Ajore, R., Niroula, A., Pertesi, M., Cafaro, C., Thodberg, M., Went, M., Bao, E. L., Duran-Lozano, L., Lopez de Lapuente Portilla, A., Olafsdottir, T., Ugidos-Damboriena, N., Magnusson, O., Samur, M., Lareau, C. A., Halldorsson, G. H., Thorleifsson, G., Norddahl, G. L., Gunnarsdottir, K., Forsti, A., Goldschmidt, H., Hemminki, K., van Rhee, F., Kimber, S., Sperling, A. S., Kaiser, M., Anderson, K., Jonsdottir, I., Munshi, N., Rafnar, T., Waage, A., Weinhold, N., Thorsteinsdottir, U., Sankaran, V. G., Stefansson, K., Houlston, R., Nilsson, B. 1800; 13 (1): 151

  Abstract

  Thousands of non-coding variants have been associated with increased risk of human diseases, yet the causal variants and their mechanisms-of-action remain obscure. In an integrative study combining massively parallel reporter assays (MPRA), expression analyses (eQTL, meQTL, PCHiC) and chromatin accessibility analyses in primary cells (caQTL), we investigate 1,039 variants associated with multiple myeloma (MM). We demonstrate that MM susceptibility is mediated by gene-regulatory changes in plasma cells and B-cells, and identify putative causal variants at six risk loci (SMARCD3, WAC, ELL2, CDCA7L, CEP120, and PREX1). Notably, three of these variants co-localize with significant plasma cell caQTLs, signaling the presence of causal activity at these precise genomic positions in an endogenous chromosomal context in vivo. Our results provide a systematic functional dissection of risk loci for a hematologic malignancy.

  View details for DOI 10.1038/s41467-021-27666-x

  View details for PubMedID 35013207

• Development of PDE6D and CK1alpha Degraders through Chemical Derivatization of FPFT-2216. Journal of medicinal chemistry Teng, M., Lu, W., Donovan, K. A., Sun, J., Krupnick, N. M., Nowak, R. P., Li, Y., Sperling, A. S., Zhang, T., Ebert, B. L., Fischer, E. S., Gray, N. S. 1800

  Abstract

  Immunomodulatory drugs are a class of drugs approved for the treatment of multiple myeloma. These compounds exert their clinical effects by inducing interactions between the CRL4CRBN E3 ubiquitin ligase and a C2H2 zinc finger degron motif, resulting in degradation of degron-containing targets. However, although many cellular proteins feature the degron motif, only a subset of those are degradable via this strategy. Here, we demonstrated that FPFT-2216, a previously reported "molecular glue" compound, degrades PDE6D, in addition to IKZF1, IKZF3, and CK1alpha. We used FPFT-2216 as a starting point for a focused medicinal chemistry campaign and developed TMX-4100 and TMX-4116, which exhibit greater selectivity for degrading PDE6D and CK1alpha, respectively. We also showed that the region in PDE6D that interacts with the FPFT-2216 derivatives is not the previously pursued prenyl-binding pocket. Moreover, we found that PDE6D depletion by FPFT-2216 does not impede the growth of KRASG12C-dependent MIA PaCa-2 cells, highlighting the challenges of drugging PDE6D-KRAS. Taken together, the approach we described here represents a general scheme to rapidly develop selective degraders by reprogramming E3 ubiquitin ligase substrate specificity.

  View details for DOI 10.1021/acs.jmedchem.1c01832

  View details for PubMedID 34965125

• Modeling and targeting of erythroleukemia by hematopoietic genome editing. Blood Iacobucci, I., Qu, C., Varotto, E., Janke, L. J., Yang, X., Seth, A., Shelat, A., Friske, J. D., Fukano, R., Yu, J., Freeman, B. B., Kennedy, J. A., Sperling, A. S., Zheng, R., Wang, Y., Jogiraju, H., Dickerson, K. M., Payne-Turner, D., Morris, S. M., Hollis, E. S., Ghosn, N., Haggard, G. E., Lindsley, R. C., Ebert, B. L., Mullighan, C. G. 2021; 137 (12): 1628-1640

  Abstract

  Acute erythroid leukemia (AEL) is characterized by a distinct morphology, mutational spectrum, lack of preclinical models, and poor prognosis. Here, using multiplexed genome editing of mouse hematopoietic stem and progenitor cells and transplant assays, we developed preclinical models of AEL and non-erythroid acute leukemia and describe the central role of mutational cooperativity in determining leukemia lineage. Different combination of mutations in Trp53, Bcor, Dnmt3a, Rb1, and Nfix resulted in the development of leukemia with an erythroid phenotype, accompanied by the acquisition of alterations in signaling and transcription factor genes that recapitulate human AEL by cross-species genomic analysis. Clonal expansion during tumor evolution was driven by mutational cooccurrence, with clones harboring a higher number of founder and secondary lesions (eg, mutations in signaling genes) showing greater evolutionary fitness. Mouse and human AEL exhibited deregulation of genes regulating erythroid development, notably Gata1, Klf1, and Nfe2, driven by the interaction of mutations of the epigenetic modifiers Dnmt3a and Tet2 that perturbed methylation and thus expression of lineage-specific transcription factors. The established mouse leukemias were used as a platform for drug screening. Drug sensitivity was associated with the leukemia genotype, with the poly (ADP-ribose) polymerase inhibitor talazoparib and the demethylating agent decitabine efficacious in Trp53/Bcor-mutant AEL, CDK7/9 inhibitors in Trp53/Bcor/Dnmt3a-mutant AEL, and gemcitabine and bromodomain inhibitors in NUP98-KDM5A leukemia. In conclusion, combinatorial genome editing has shown the interplay of founding and secondary genetic alterations in phenotype and clonal evolution, epigenetic regulation of lineage-specific transcription factors, and therapeutic tractability in erythroid leukemogenesis.

  View details for DOI 10.1182/blood.2020009103

  View details for PubMedID 33512458

• A deep molecular response of splenic marginal zone lymphoma to front-line checkpoint blockade HAEMATOLOGICA Miller, P. G., Sperling, A. S., Gibson, C. J., Pozdnyakova, O., Wong, W. J., Manos, M. P., Buchbinder, E. I., Hodi, F., Ebert, B. L., Davids, M. S. 2021; 106 (2): 651-654

  View details for DOI 10.3324/haematol.2020.258426

  View details for Web of Science ID 000615767000021

  View details for PubMedID 33054119

  View details for PubMedCentralID PMC7849751

• Reversible ON- and OFF-switch chimeric antigen receptors controlled by lenalidomide SCIENCE TRANSLATIONAL MEDICINE Jan, M., Scarfo, I., Larson, R. C., Walker, A., Schmidts, A., Guirguis, A. A., Gasser, J. A., Slabicki, M., Bouffard, A. A., Castano, A. P., Kann, M. C., Cabral, M. L., Tepper, A., Grinshpun, D. E., Sperling, A. S., Kyung, T., Sievers, Q. L., Birnbaum, M. E., Maus, M., Ebert, B. L. 2021; 13 (575)

  Abstract

  Cell-based therapies are emerging as effective agents against cancer and other diseases. As autonomous "living drugs," these therapies lack precise control. Chimeric antigen receptor (CAR) T cells effectively target hematologic malignancies but can proliferate rapidly and cause toxicity. We developed ON and OFF switches for CAR T cells using the clinically approved drug lenalidomide, which mediates the proteasomal degradation of several target proteins by inducing interactions between the CRL4CRBN E3 ubiquitin ligase and a C2H2 zinc finger degron motif. We performed a systematic screen to identify "super-degron" tags with enhanced sensitivity to lenalidomide-induced degradation and used these degradable tags to generate OFF-switch degradable CARs. To create an ON switch, we engineered a lenalidomide-inducible dimerization system and developed split CARs that required both lenalidomide and target antigen for activation. Subtherapeutic lenalidomide concentrations controlled the effector functions of ON- and OFF-switch CAR T cells. In vivo, ON-switch split CARs demonstrated lenalidomide-dependent antitumor activity, and OFF-switch degradable CARs were depleted by drug treatment to limit inflammatory cytokine production while retaining antitumor efficacy. Together, the data showed that these lenalidomide-gated switches are rapid, reversible, and clinically suitable systems to control transgene function in diverse gene- and cell-based therapies.

  View details for DOI 10.1126/scitranslmed.abb6295

  View details for Web of Science ID 000608364300005

  View details for PubMedID 33408186

  View details for PubMedCentralID PMC8045771

• Association of Clonal Hematopoiesis with Chronic Obstructive Pulmonary Disease. Blood Miller, P., Qiao, D., Rojas-Quintero, J., Honigberg, M. C., Sperling, A. S., Gibson, C. J., Bick, A. G., Niroula, A., McConkey, M. E., Sandoval, B., Miller, B., Shi, W., Viswanathan, K., Leventhal, M. J., Werner, L., Moll, M., Cade, B., Barr, R. G., Correa, A., Cupples, L. A., Gharib, S. A., Jain, D., Gogarten, S., Lange, L., London, S., Manichaikul, A., O'Connor, G., Oelsner, E., Redline, S., Rich, S. S., Rotter, J. I., Ramachandran, V., Yu, B., Sholl, L. M., Neuberg, D., Jaiswal, S., Levy, B., Owen, C., Natarajan, P., Silverman, E. K., van Galen, P., Tesfaigzi, Y., Cho, M., Ebert, B. L. 2021

  Abstract

  Chronic obstructive pulmonary disease (COPD) is associated with age and smoking, but other determinants of the disease are incompletely understood. Clonal hematopoiesis of indeterminate potential (CHIP) is a common, age-related state in which somatic mutations in clonal blood populations induce aberrant inflammatory responses. Patients with CHIP have an elevated risk for cardiovascular disease, but the association with COPD remains unclear. We analyzed whole-genome and exome sequencing data to detect CHIP in 48,835 subjects, of whom 8,444 had moderate-to-very-severe COPD, from four separate cohorts with COPD phenotyping and smoking history. We measured emphysema in murine models in which Tet2 was deleted in hematopoietic cells. In COPDGene, individuals with CHIP had a risk of moderate-to-severe and severe or very severe COPD 1.6 and 2.2 times greater than non-carriers, respectively (adjusted 95% confidence intervals [CI], 1.1 to 2.2 and 1.5 to 3.2). These findings were consistent observed in three additional cohorts and meta-analyses of all subjects. CHIP was also associated with decreased FEV1% predicted in COPDGene (mean between group difference -5.7%; adjusted 95% CI, -8.8 to -2.6), a finding replicated in additional cohorts. Smoke exposure was associated with a small but significant increased risk of having CHIP (OR 1.03 per ten pack-years, 95% CI 1.01-1.05) in the meta-analysis of all subjects. Inactivation of Tet2 in mouse hematopoietic cells exacerbated emphysema development and inflammation in cigarette smoke exposure models. Somatic mutations in blood cells are associated with the development and severity of COPD, independent of age and cumulative smoke exposure.

  View details for DOI 10.1182/blood.2021013531

  View details for PubMedID 34855941

• Contribution of clonal hematopoiesis to adult-onset hemophagocytic lymphohistiocytosis BLOOD Miller, P. G., Sperling, A. S., Gibson, C. J., Viswanathan, K., Castellano, C., McConkey, M., Ceremsak, J., Taylor, M. S., Birndt, S., Perner, F., Arnason, J., Agrawal, M., Schram, A. M., Nikiforow, S., Pihan, G., Hasserjian, R. P., Aster, J. C., La Rosee, P., Morgan, E. A., Berliner, N., Ebert, B. L. 2020; 136 (26): 3051-3055

  Abstract

  Adult-onset hemophagocytic lymphohistiocytosis (HLH) is a rare, life-threatening disease of immune hyperactivation. Unlike pediatric HLH, adult HLH is rarely driven by germline genetic variants. Although numerous precipitating etiologies have been identified, the reason that HLH occurs in only a subset of individuals and how other factors contribute to the disease remains unknown. We hypothesized that clonal hematopoiesis (CH), a state in which somatic mutations in blood cells cause an expanded population of mutant hematopoietic cells and drive an aberrant inflammatory state, could contribute to adult-onset HLH. In a highly annotated cohort of older adults with HLH we found that CH was more prevalent than in control cohorts. Using the adult-onset HLH mouse model in which repeated treatments of the TLR9 agonist, ODN1826, was delivered to the mouse, we observed that macrophages carrying mutations in Tet2, one of the most commonly mutated genes in CH, have an enhanced inflammatory response to TLR9 agonism. Finally, mice carrying Tet2 mutations in the hematopoietic compartment (a common model for CH) displayed an exaggerated response to TLR9 agonism, including worse splenomegaly and anemia. Our data suggest that CH is more common in individuals with adult-onset HLH and can contribute to the pathophysiology of this disease.

  View details for DOI 10.1182/blood.2020008206

  View details for Web of Science ID 000603450500013

  View details for PubMedID 32961550

  View details for PubMedCentralID PMC7770567

• Gait speed, survival, and recommended treatment intensity in older adults with blood cancer requiring treatment CANCER Hantel, A., DuMontier, C., Odejide, O. O., Luskin, M. R., Sperling, A. S., Hshieh, T., Chen, R., Soiffer, R., Driver, J. A., Abel, G. A. 2021; 127 (6): 875-883

  Abstract

  Brief measures of physical function such as gait speed may be useful to optimize treatment intensity for older adults who have blood cancer; however, little is known about whether such assessments are already captured within oncologists' "gestalt" assessments.Gait speed was assessed in 782 patients ≥75 years of age who had blood cancer, with results reported to providers after treatment decisions were made; 408 patients required treatment when different intensities were available per National Comprehensive Cancer Network (NCCN) guidelines. We performed structured abstractions of treatment intensity recommendations into standard intensity, reduced intensity, or supportive care, based on NCCN guidelines. We modeled gait speed and survival using Cox regression and performed ordinal logistic regression to assess predictors of NCCN-based categorizations of oncologists' treatment intensity recommendations, including gait speed.The median survival by gait speed category was 10.8 months (<0.4 m/s), 18.6 months (0.4-0.6 m/s), 34.0 months (0.6-0.8 m/s), and unreached (>0.8 m/s). Univariable hazard ratios (HRs) for death increased for each lower category compared with ≥0.8 m/s (0.6-0.8 m/s: HR, 1.76; 0.4-0.6 m/s: HR, 2.30; <0.4 m/s: HR, 3.31). Gait speed predicted survival in multivariable Cox regression (all P < .05). In multivariable models including age, sex, and Eastern Cooperative Oncology Group performance status, gait speed did not predict oncologists' recommended treatment intensity (all P > .05) and did not add to a base model predicting recommended treatment intensity.In older adults with blood cancer who presented for treatment, gait speed predicted survival but not treatment intensity recommendation. Incorporating gait speed into decision making may improve optimal treatment selection.

  View details for DOI 10.1002/cncr.33344

  View details for Web of Science ID 000592028300001

  View details for PubMedID 33237587

  View details for PubMedCentralID PMC7946649

• Small-molecule-induced polymerization triggers degradation of BCL6 NATURE Slabicki, M., Yoon, H., Koeppel, J., Nitsch, L., Roy Burman, S. S., Di Genua, C., Donovan, K. A., Sperling, A. S., Hunkeler, M., Tsai, J. M., Sharma, R., Guirguis, A., Zou, C., Chudasama, P., Gasser, J. A., Miller, P. G., Scholl, C., Froehling, S., Nowak, R. P., Fischer, E. S., Ebert, B. L. 2020; 588 (7836): 164-+

  Abstract

  Effective and sustained inhibition of non-enzymatic oncogenic driver proteins is a major pharmacological challenge. The clinical success of thalidomide analogues demonstrates the therapeutic efficacy of drug-induced degradation of transcription factors and other cancer targets1-3, but a substantial subset of proteins are resistant to targeted degradation using existing approaches4,5. Here we report an alternative mechanism of targeted protein degradation, in which a small molecule induces the highly specific, reversible polymerization of a target protein, followed by its sequestration into cellular foci and subsequent degradation. BI-3802 is a small molecule that binds to the Broad-complex, Tramtrack and Bric-à-brac (BTB) domain of the oncogenic transcription factor B cell lymphoma 6 (BCL6) and leads to the proteasomal degradation of BCL66. We use cryo-electron microscopy to reveal how the solvent-exposed moiety of a BCL6-binding molecule contributes to a composite ligand-protein surface that engages BCL6 homodimers to form a supramolecular structure. Drug-induced formation of BCL6 filaments facilitates ubiquitination by the SIAH1 E3 ubiquitin ligase. Our findings demonstrate that a small molecule such as BI-3802 can induce polymerization coupled to highly specific protein degradation, which in the case of BCL6 leads to increased pharmacological activity compared to the effects induced by other BCL6 inhibitors. These findings open new avenues for the development of therapeutic agents and synthetic biology.

  View details for DOI 10.1038/s41586-020-2925-1

  View details for Web of Science ID 000591047800004

  View details for PubMedID 33208943

  View details for PubMedCentralID PMC7816212

• Fitness Landscape of Clonal Hematopoiesis Under Selective Pressure of Immune Checkpoint Blockade JCO PRECISION ONCOLOGY Miller, P. G., Gibson, C. J., Mehta, A., Sperling, A. S., Frederick, D. T., Manos, M. P., Miao, B., Hacohen, N., Hodi, F., Boland, G. M., Ebert, B. L. 2020; 4: 1027-1033

  Abstract

  Conventional cytotoxic therapies increase the risk of clonal hematopoiesis and select for TP53-mutant clones, which carry a high risk for transformation to therapy-related myelodysplastic neoplasms. In contrast, the effect of immune checkpoint blockade (ICB) on clonal hematopoiesis is unknown.Paired peripheral-blood samples taken before and after treatment with ICB were obtained for 91 patients with either cutaneous melanoma or basal cell carcinoma. Error-corrected sequencing of a targeted panel of genes recurrently mutated in clonal hematopoiesis was performed on peripheral-blood genomic DNA.The average interval between acquisition of the paired samples was 180 days. Forty-one percent of the patients had clonal hematopoiesis at a variant allele frequency (VAF) > 0.01 in the pretreatment sample. There was near-complete agreement in the distribution and burden of clonal hematopoiesis mutations in the paired blood samples, with 87 of 88 mutations identified across the cohort present in paired samples, regardless of the duration between sample collection. The VAF in the paired samples also showed a high correlation, with an R 2 = 0.95 (P < .0001). In contrast to cytotoxic therapy, exposure to ICB did not lead to selection of TP53- or PPM1D-mutant clones. However, consistent with the known effects of DNA-damaging therapy, we identified one patient who had eight unique TP53 mutations in the posttreatment blood sample after receiving two courses of radiation therapy.There was no expansion of hematopoietic clones or selection for clones at high risk for malignant transformation in patients who received ICB, observations that warrant further validation in larger cohorts. These findings highlight an important difference between ICB and conventional cytotoxic therapies and their respective impacts on premalignant genetic lesions.

  View details for DOI 10.1200/PO.20.00186

  View details for Web of Science ID 000615676400001

  View details for PubMedID 33015529

  View details for PubMedCentralID PMC7529531

• The CDK inhibitor CR8 acts as a molecular glue degrader that depletes cyclin K NATURE Slabicki, M., Kozicka, Z., Petzold, G., Li, Y., Manojkumar, M., Bunker, R. D., Donovan, K. A., Sievers, Q. L., Koeppel, J., Suchyta, D., Sperling, A. S., Fink, E. C., Gasser, J. A., Wang, L. R., Corsello, S. M., Sellar, R. S., Jan, M., Gillingham, D., Scholl, C., Frohling, S., Golub, T. R., Fischer, E. S., Thoma, N. H., Ebert, B. L. 2020; 585 (7824): 293-+

  Abstract

  Molecular glue compounds induce protein-protein interactions that, in the context of a ubiquitin ligase, lead to protein degradation1. Unlike traditional enzyme inhibitors, these molecular glue degraders act substoichiometrically to catalyse the rapid depletion of previously inaccessible targets2. They are clinically effective and highly sought-after, but have thus far only been discovered serendipitously. Here, through systematically mining databases for correlations between the cytotoxicity of 4,518 clinical and preclinical small molecules and the expression levels of E3 ligase components across hundreds of human cancer cell lines3-5, we identify CR8-a cyclin-dependent kinase (CDK) inhibitor6-as a compound that acts as a molecular glue degrader. The CDK-bound form of CR8 has a solvent-exposed pyridyl moiety that induces the formation of a complex between CDK12-cyclin K and the CUL4 adaptor protein DDB1, bypassing the requirement for a substrate receptor and presenting cyclin K for ubiquitination and degradation. Our studies demonstrate that chemical alteration of surface-exposed moieties can confer gain-of-function glue properties to an inhibitor, and we propose this as a broader strategy through which target-binding molecules could be converted into molecular glues.

  View details for DOI 10.1038/s41586-020-2374-x

  View details for Web of Science ID 000537663800004

  View details for PubMedID 32494016

  View details for PubMedCentralID PMC7486275

• Identification of germline variants in adults with hemophagocytic lymphohistiocytosis BLOOD ADVANCES Miller, P. G., Niroula, A., Ceremsak, J. J., Gibson, C. J., Taylor, M. S., Birndt, S., Perner, F., Arnason, J., Sperling, A. S., Agrawal, M., Schram, A. M., Nikiforow, S., Pihan, G., Hasserjian, R. P., Aster, J. C., La Rosee, P., Morgan, E. A., Berliner, N., Ebert, B. L. 2020; 4 (5): 925-929

  Abstract

  Some germline variants are predicted to disrupt protein function in HLH-associated genes. Such variants are neither enriched in adult-onset HLH nor associated with specific clinical or laboratory features of HLH.

  View details for DOI 10.1182/bloodadvances.2019001272

  View details for Web of Science ID 000519811200016

  View details for PubMedID 32150605

  View details for PubMedCentralID PMC7065469

• CHIPing Away at Breast Cancer JNCI-JOURNAL OF THE NATIONAL CANCER INSTITUTE Sperling, A. S., Ebert, B. L. 2020; 112 (1): 10-11

  View details for DOI 10.1093/jnci/djz158

  View details for Web of Science ID 000510453200005

  View details for PubMedID 31504659

• Increased mitochondrial apoptotic priming with targeted therapy predicts clinical response to re-induction chemotherapy. American journal of hematology Garcia, J. S., Bhatt, S., Fell, G., Sperling, A. S., Burgess, M., Keshishian, H., Yilma, B., Brunner, A., Neuberg, D., Carr, S. A., Ebert, B. L., Ballen, K., Stone, R. M., DeAngelo, D. J., Medeiros, B. C., Letai, A. 2019

  Abstract

  Most patients with relapsed or refractory (R/R) acute myeloid leukemia (AML) do not benefit from current re-induction or approved targeted therapies. In the absence of targetable genetic mutations, there is minimal guidance on optimal treatment selection particularly in the R/R setting highlighting an unmet need for clinically useful functional biomarkers. Blood and bone marrow samples from patients treated on two clinical trials testing the combination of lenalidomide (LEN) and MEC (mitoxantrone, etoposide, and cytarabine) chemotherapy in R/R AML patients were available to test the clinical utility of the mitochondrial apoptotic BH3 and dynamic BH3 profiling (DBP) assays in predicting response, as there was no clear genetic biomarker identifying responders. To test whether LEN-induced mitochondrial priming predicted clinical response to LEN-MEC therapy, we performed DBP on patient myeloblasts and found that short-term ex vivo treatment with lenalidomide discriminated clinical responders from non-responders based on drug-induced change in priming (delta priming). Using paired patient samples collected before and after clinical LEN treatment (prior to MEC dosing), we confirmed LEN-induced increased apoptotic priming in vivo, suggesting LEN enhanced vulnerability of myeloblasts to cytotoxic MEC chemotherapy. This is the first study demonstrating the potential role of DBP in predicting clinical response to a combination regimen. Our findings demonstrate that functional properties of relapsed AML can identify active therapies. This article is protected by copyright. All rights reserved.

  View details for DOI 10.1002/ajh.25692

  View details for PubMedID 31804723

• Plasmacytoma presenting as jugular foramen tumor in a young woman with multiple myeloma AMERICAN JOURNAL OF HEMATOLOGY How, J., Johnson, P., Corrales, C., Wright, K., Justicz, N., Chen, J. X., Sperling, A. S., Connell, N. T. 2019; 94 (6): 728-732

  View details for DOI 10.1002/ajh.25477

  View details for Web of Science ID 000467587800026

  View details for PubMedID 30916796

• Widespread intronic polyadenylation diversifies immune cell transcriptomes NATURE COMMUNICATIONS Singh, I., Lee, S., Sperling, A. S., Samur, M. K., Tai, Y., Fulciniti, M., Munshi, N. C., Mayr, C., Leslie, C. S. 2018; 9: 1716

  Abstract

  Alternative cleavage and polyadenylation (ApA) is known to alter untranslated region (3'UTR) length but can also recognize intronic polyadenylation (IpA) signals to generate transcripts that lose part or all of the coding region. We analyzed 46 3'-seq and RNA-seq profiles from normal human tissues, primary immune cells, and multiple myeloma (MM) samples and created an atlas of 4927 high-confidence IpA events represented in these cell types. IpA isoforms are widely expressed in immune cells, differentially used during B-cell development or in different cellular environments, and can generate truncated proteins lacking C-terminal functional domains. This can mimic ectodomain shedding through loss of transmembrane domains or alter the binding specificity of proteins with DNA-binding or protein-protein interaction domains. MM cells display a striking loss of IpA isoforms expressed in plasma cells, associated with shorter progression-free survival and impacting key genes in MM biology and response to lenalidomide.

  View details for DOI 10.1038/s41467-018-04112-z

  View details for Web of Science ID 000431113000004

  View details for PubMedID 29712909

  View details for PubMedCentralID PMC5928244

• A phase I study of lenalidomide plus chemotherapy with mitoxantrone, etoposide, and cytarabine for the reinduction of patients with acute myeloid leukemia AMERICAN JOURNAL OF HEMATOLOGY DeAngelo, D. J., Brunner, A. M., Werner, L., Avigan, D., Fathi, A. T., Sperling, A. S., Washington, A., Stroopinsky, D., Rosenblatt, J., McMasters, M., Luptakova, K., Wadleigh, M., Steensma, D. P., Hobbs, G. S., Attar, E. C., Amrein, P. C., Ebert, B. L., Stone, R. M., Ballen, K. K. 2018; 93 (2): 254-261

  Abstract

  Patients with relapsed AML have a poor prognosis and limited responses to standard chemotherapy. Lenalidomide is an immunomodulatory drug that may modulate anti-tumor immunity. We performed a study to evaluate the safety and tolerability of lenalidomide with mitoxantrone, etoposide and cytarabine (MEC) in relapsed/refractory AML. Adult patients with relapsed/refractory AML were eligible for this phase I dose-escalation study. We enrolled 35 patients using a "3 + 3" design, with a 10 patient expansion cohort at the maximum tolerated dose (MTD). Lenalidomide was initially given days 1-14 and MEC days 4-8; due to delayed count recovery, the protocol was amended to administer lenalidomide days 1-10. The dose of lenalidomide was then escalated starting at 5 mg/d (5-10-25-50). The primary objective was tolerability and MTD determination, with secondary outcomes including overall survival (OS). The MTD of lenalidomide combined with MEC was 50 mg/d days 1-10. Among the 35 enrolled patients, 12 achieved complete remission (CR) (34%, 90%CI 21-50%); 30-day mortality was 6% and 60-day mortality 13%. The median OS for all patients was 11.5 months. Among 17 patients treated at the MTD, 7 attained CR (41%); the median OS was not reached while 12-month OS was 61%. Following therapy with MEC and lenalidomide, patient CD4+ and CD8+ T-cells demonstrated increased inflammatory responses to autologous tumor lysate. The combination of MEC and lenalidomide is tolerable with an RP2D of lenalidomide 50 mg/d days 1-10, yielding encouraging response rates. Further studies are planned to explore the potential immunomodulatory effect of lenalidomide and MEC.

  View details for DOI 10.1002/ajh.24968

  View details for Web of Science ID 000419455900022

  View details for PubMedID 29119643

  View details for PubMedCentralID PMC6290473

• Prevalent premalignancy BLOOD Sperling, A. S., Ebert, B. L. 2017; 130 (6): 695-696

  View details for DOI 10.1182/blood-2017-05-786210

  View details for Web of Science ID 000407393300002

  View details for PubMedID 28798057

• Factoring in the missing link AMERICAN JOURNAL OF HEMATOLOGY Sethi, N., Manesh, R., Sperling, A., Bresler, S. C., Connell, N. T., Tierney, L. M. 2017; 92 (1): 110-113

  View details for DOI 10.1002/ajh.24501

  View details for Web of Science ID 000393425300027

  View details for PubMedID 27486087

  View details for PubMedCentralID PMC7261369

• A DNA target-enrichment approach to detect mutations, copy number changes and immunoglobulin translocations in multiple myeloma BLOOD CANCER JOURNAL Bolli, N., Li, Y., Sathiaseelan, V., Raine, K., Jones, D., Ganly, P., Cocito, F., Bignell, G., Chapman, M. A., Sperling, A. S., Anderson, K. C., Avet-Loiseau, H., Minvielle, S., Campbell, P. J., Munshi, N. C. 2016; 6: e467

  Abstract

  Genomic lesions are not investigated during routine diagnostic workup for multiple myeloma (MM). Cytogenetic studies are performed to assess prognosis but with limited impact on therapeutic decisions. Recently, several recurrently mutated genes have been described, but their clinical value remains to be defined. Therefore, clinical-grade strategies to investigate the genomic landscape of myeloma samples are needed to integrate new and old prognostic markers. We developed a target-enrichment strategy followed by next-generation sequencing (NGS) to streamline simultaneous analysis of gene mutations, copy number changes and immunoglobulin heavy chain (IGH) translocations in MM in a high-throughput manner, and validated it in a panel of cell lines. We identified 548 likely oncogenic mutations in 182 genes. By integrating published data sets of NGS in MM, we retrieved a list of genes with significant relevance to myeloma and found that the mutational spectrum of primary samples and MM cell lines is partially overlapping. Gains and losses of chromosomes, chromosomal segments and gene loci were identified with accuracy comparable to conventional arrays, allowing identification of lesions with known prognostic significance. Furthermore, we identified IGH translocations with high positive and negative predictive value. Our approach could allow the identification of novel biomarkers with clinical relevance in myeloma.

  View details for DOI 10.1038/bcj.2016.72

  View details for Web of Science ID 000391802800003

  View details for PubMedID 27588520

  View details for PubMedCentralID PMC5056967

• Heterogeneity of genomic evolution and mutational profiles in multiple myeloma NATURE COMMUNICATIONS Bolli, N., Avet-Loiseau, H., Wedge, D. C., Van Loo, P., Alexandrov, L. B., Martincorena, I., Dawson, K. J., Iorio, F., Nik-Zainal, S., Bignell, G. R., Hinton, J. W., Li, Y., Tubio, J. M. C., McLaren, S., Meara, S., Butler, A. P., Teague, J. W., Mudie, L., Anderson, E., Rashid, N., Tai, Y., Shammas, M. A., Sperling, A. S., Fulciniti, M., Richardson, P. G., Parmigiani, G., Magrangeas, F., Minvielle, S., Moreau, P., Attal, M., Facon, T., Futreal, P., Anderson, K. C., Campbell, P. J., Munshi, N. C. 2014; 5: 2997

  Abstract

  Multiple myeloma is an incurable plasma cell malignancy with a complex and incompletely understood molecular pathogenesis. Here we use whole-exome sequencing, copy-number profiling and cytogenetics to analyse 84 myeloma samples. Most cases have a complex subclonal structure and show clusters of subclonal variants, including subclonal driver mutations. Serial sampling reveals diverse patterns of clonal evolution, including linear evolution, differential clonal response and branching evolution. Diverse processes contribute to the mutational repertoire, including kataegis and somatic hypermutation, and their relative contribution changes over time. We find heterogeneity of mutational spectrum across samples, with few recurrent genes. We identify new candidate genes, including truncations of SP140, LTB, ROBO1 and clustered missense mutations in EGR1. The myeloma genome is heterogeneous across the cohort, and exhibits diversity in clonal admixture and in dynamics of evolution, which may impact prognostic stratification, therapeutic approaches and assessment of disease response to treatment.

  View details for DOI 10.1038/ncomms3997

  View details for Web of Science ID 000331081900001

  View details for PubMedID 24429703

  View details for PubMedCentralID PMC3905727

• γH2A is a component of yeast heterochromatin required for telomere elongation CELL CYCLE Kitada, T., Schleker, T., Sperling, A. S., Xie, W., Gasser, S. M., Grunstein, M. 2011; 10 (2): 293-300

  Abstract

  Histones of heterochromatin are deacetylated in yeast and methylated in more complex eukaryotes to regulate heterochromatin structure and gene silencing. Here, we report that histone H2A phosphorylated at serine 129 (γH2A) in Saccharomyces cerevisiae is a conceptually new type of heterochromatin modification that functions downstream of silent chromatin assembly. We show that γH2A is enriched throughout yeast telomeric and silent mating locus (HM) heterochromatin where γH2A results from the action of kinases Tel1 and Mec1. Interestingly, mutation of γH2A has no apparent effect on the binding of Sir (silent information regulator) complex or on gene silencing. In contrast, deletion of SIR3 abolishes the formation of γH2A at heterochromatin. To address the function of γH2A, we used a Δrif1 mutant strain in which telomeres are excessively elongated to show that γH2A is required for the optimal recruitment of Cdc13, a regulator of telomere elongation, and for telomere elongation itself. Thus, a histone modification that parallels Sir3 protein binding is shown here to be dispensable for the formation of a silent structure but is important for a crucial heterochromatin-specific downstream function in telomere homeostasis.

  View details for DOI 10.4161/cc.10.2.14536

  View details for Web of Science ID 000286461600025

  View details for PubMedID 21212735

  View details for PubMedCentralID PMC3033431

• Histone H3 Lysine 56 Acetylation Is Linked to the Core Transcriptional Network in Human Embryonic Stem Cells MOLECULAR CELL Xie, W., Song, C., Young, N. L., Sperling, A. S., Xu, F., Sridharan, R., Conway, A. E., Garcia, B. A., Plath, K., Clark, A. T., Grunstein, M. 2009; 33 (4): 417-427

  Abstract

  Lysine 56 acetylation in the helical core of histone H3 opens yeast chromatin and enables histone gene transcription, DNA replication, and DNA repair and prevents epigenetic silencing. While K56Ac is globally abundant in yeast and flies, its presence has been uncertain in mammals. We show here using mass spectrometry and genome-wide analyses that K56Ac is present in human embryonic stem cells (hESCs), overlapping strongly at active and inactive promoters with the binding of the key regulators of pluripotency, NANOG, SOX2, and OCT4. This includes also the canonical histone gene promoters and those for the hESC-specific microRNAs. K56Ac then relocates to developmental genes upon cellular differentiation. Thus the K56Ac state more accurately reflects the epigenetic differences between hESCs and somatic cells than other active histone marks such as H3 K4 trimethylation and K9 acetylation. These results suggest that K56Ac is involved in the human core transcriptional network of pluripotency.

  View details for DOI 10.1016/j.molcel.2009.02.004

  View details for Web of Science ID 000263810800004

  View details for PubMedID 19250903

  View details for PubMedCentralID PMC2671231

• Genetic Identification of a Network of Factors that Functionally Interact with the Nucleosome Remodeling ATPase <i>ISWI</i> PLOS GENETICS Burgio, G., La Rocca, G., Sala, A., Arancio, W., Di Gesu, D., Collesano, M., Sperling, A. S., Armstrong, J. A., van Heeringen, S. J., Logie, C., Tamkun, J. W., Corona, D. F. V. 2008; 4 (6): e1000089

  Abstract

  Nucleosome remodeling and covalent modifications of histones play fundamental roles in chromatin structure and function. However, much remains to be learned about how the action of ATP-dependent chromatin remodeling factors and histone-modifying enzymes is coordinated to modulate chromatin organization and transcription. The evolutionarily conserved ATP-dependent chromatin-remodeling factor ISWI plays essential roles in chromosome organization, DNA replication, and transcription regulation. To gain insight into regulation and mechanism of action of ISWI, we conducted an unbiased genetic screen to identify factors with which it interacts in vivo. We found that ISWI interacts with a network of factors that escaped detection in previous biochemical analyses, including the Sin3A gene. The Sin3A protein and the histone deacetylase Rpd3 are part of a conserved histone deacetylase complex involved in transcriptional repression. ISWI and the Sin3A/Rpd3 complex co-localize at specific chromosome domains. Loss of ISWI activity causes a reduction in the binding of the Sin3A/Rpd3 complex to chromatin. Biochemical analysis showed that the ISWI physically interacts with the histone deacetylase activity of the Sin3A/Rpd3 complex. Consistent with these findings, the acetylation of histone H4 is altered when ISWI activity is perturbed in vivo. These findings suggest that ISWI associates with the Sin3A/Rpd3 complex to support its function in vivo.

  View details for DOI 10.1371/journal.pgen.1000089

  View details for Web of Science ID 000260410300025

  View details for PubMedID 18535655

  View details for PubMedCentralID PMC2390755

• The Drosophila BRM complex facilitates global transcription by RNA polymerase II EMBO JOURNAL Armstrong, J. A., Papoulas, O., Daubresse, G., Sperling, A. S., Lis, J. T., Scott, M. P., Tamkun, J. W. 2002; 21 (19): 5245-5254

  Abstract

  Drosophila brahma (brm) encodes the ATPase subunit of a 2 MDa complex that is related to yeast SWI/SNF and other chromatin-remodeling complexes. BRM was identified as a transcriptional activator of Hox genes required for the specification of body segment identities. To clarify the role of the BRM complex in the transcription of other genes, we examined its distribution on larval salivary gland polytene chromosomes. The BRM complex is associated with nearly all transcriptionally active chromatin in a pattern that is generally non-overlapping with that of Polycomb, a repressor of Hox gene transcription. Reduction of BRM function dramatically reduces the association of RNA polymerase II with salivary gland chromosomes. A few genes, such as induced heat shock loci, are not associated with the BRM complex; transcription of these genes is not compromised by loss of BRM function. The distribution of the BRM complex thus correlates with a dependence on BRM for gene activity. These data suggest that the chromatin remodeling activity of the BRM complex plays a general role in facilitating transcription by RNA polymerase II.

  View details for Web of Science ID 000178502600024

  View details for PubMedID 12356740

  View details for PubMedCentralID PMC129039