Dr. Justin Annes specializes in the treatment of hereditary endocrine disorders with particular focus on neuroendocrine-related conditions.
He developed the Stanford Endocrine Genetics Clinic in 2012 which is part of the interdisciplinary Stanford Hypertension Center and Stanford Neuroendocrine Tumor Program.
He has medical practice has focused on hereditary endocrine disease since 2008.

Clinical Focus

  • Pheochromocytoma and Paraganglioma
  • Multiple Endocrine Neoplasia
  • Hereditary Endocrine Disorders
  • Endocrinology
  • Diabetes and Metabolism

Honors & Awards

  • Stanford Cancer Center Innovation Award, Stanford University (2018)
  • Stanford Diabetes Research Center, Training Outreach Program (PI), Stanford University (2017-)
  • Translation Research Innovation Award, The Friedenrich BII Diabetes Fund (2016-2017)
  • R01: The Role of Adenosine Kinase in Controlling Beta-Cell Regeneration, NIH NIDDK (2015-20)
  • The Best of Basic Research, Endocrine Society (2014)
  • Cellome Award for "Best High-Content Screening Publication in 2012", Thermo Fisher Scientific (2013)
  • Hoopes Mentorship Award, Harvard University (2012)
  • Mentored Clinical Scientist Research Career Development Award, NIH (2009-2014)
  • AOA, Delta Chapter, NYU Medical School (2004)
  • Medical Scientist Training Award, NIH (1996-2002)

Boards, Advisory Committees, Professional Organizations

  • Medical Advisory Board Member, Pheo Para Alliance (2018 - Present)
  • Admissions Committee, Stanford MSTP (2013 - Present)

Professional Education

  • Medical Education: NYU Grossman School of Medicine (2004) NY
  • Residency: Childrens Hospital Harvard Medical School Medical Genetics Residency (2009) MA
  • Residency: Brigham and Women's Hospital Internal Medicine Residency (2009) MA
  • Internship: Brigham and Women's Hospital Internal Medicine Residency (2005) MA
  • Fellowship, Brigham and Women's Hospital / Harvard Medical School, Clinical Genetics (2009)
  • Residency, Brigham and Women's Hospital, Internal Medicine (2009)
  • M.D., Ph.D., NYU School of Medicine, Cell Biology (2004)
  • BS, Haverford College, Molecular Biology (1996)

Current Research and Scholarly Interests

The ANNES LABORATORY of Molecular Endocrinology: Leveraging Chemical Biology to Treat Endocrine Disorders

The prevalence of diabetes is increasing at a staggering rate. By the year 2050 an astounding 25% of Americans will be diabetic. The goal of my research is to uncover therapeutic strategies to stymie the ensuing diabetes epidemic. To achieve this goal we have developed a variety of innovate experimental approaches to uncover novel approaches to curing diabetes.

(1) Beta-Cell Regeneration: Diabetes results from either an absolute or relative deficiency in insulin production. Our therapeutic strategy is to stimulate the regeneration of insulin-producing beta-cells to enhance an individual’s insulin secretion capacity. We have developed a unique high-throughput chemical screening platform which we use to identify small molecules that promote beta-cell growth. This work has led to the identification of key molecular pathways (therapeutic targets) and candidate drugs that promote the growth and regeneration of islet beta-cells. Our goal is to utilize these discoveries to treat and prevent diabetes.

(2) The Metabolic Syndrome: A major cause of the diabetes epidemic is the rise in obesity which leads to a cluster of diabetes- and cardiovascular disease-related metabolic abnormalities that shorten life expectancy. These physiologic aberrations are collectively termed the Metabolic Syndrome (MS). My laboratory has developed an original in vivo screening platform t to identify novel hormones that influence the behaviors (excess caloric consumption, deficient exercise and disrupted sleep-wake cycles) and the metabolic abnormalities caused by obesity. We aim to manipulate these hormone levels to prevent the development and detrimental consequences of the MS.

The Hereditary Paraganglioma Syndrome (hPGL) is a rare genetic cancer syndrome that is most commonly caused by a defect in mitochondrial metabolism. Our goal is to understand how altered cellular metabolism leads to the development of cancer. Although hPGL is uncommon, it serves as an excellent model for the abnormal metabolic behavior displayed by nearly all cancers. Our goal is to develop novel therapeutic strategies that target the abnormal behavior of cancer cells. In the laboratory we have developed hPGL mouse models and use high throughput chemical screening to identify the therapeutic susceptibilities that result from the abnormal metabolic behavior of cancer cells.

As a physician scientist trained in clinical genetics I have developed expertise in hereditary endocrine disorders and devoted my efforts to treating families affected by the hPGL syndrome. By leveraging our laboratory expertise in the hPGL syndrome, our care for individuals who have inherited the hPGL syndrome is at the forefront of medicine. Our goal is to translate our laboratory discoveries to the treatment of affected families.

2023-24 Courses

Stanford Advisees

Graduate and Fellowship Programs

All Publications

  • An shRNA screen in primary human beta cells identifies the serotonin 1F receptor as a negative regulator of survival during transplant. bioRxiv : the preprint server for biology Lee, R. A., Chopra, D. G., Nguyen, V., Huang, X., Zhang, Y., Shariati, K., Yiv, N., Schugar, R., Annes, J., Roth, B., Ku, G. M. 2024


    Islet transplantation can cure type 1 diabetes, but peri-transplant beta cell death limits this procedure to those with low insulin requirements. Improving human beta cell survival or proliferation may make islet transplantation a possibility for more type 1 patients. To identify novel regulators of beta cell survival and proliferation, we conducted a pooled small hairpin RNA (shRNA) screen in primary human beta cells transplanted into immunocompromised mice. shRNAs targeting several cyclin dependent kinase inhibitors were enriched after transplant. Here, we focused on the Gi/o-coupled GPCR, serotonin 1F receptor ( HTR1F, 5-HT 1F ) which our screen identified as a negative regulator of beta cell numbers after transplant. In vitro , 5-HT 1F knockdown induced human beta cell proliferation but only when combined with harmine and exendin-4. In vivo , knockdown of 5-HT 1F reduced beta cell death during transplant. To demonstrate the feasibility of targeting 5-HT 1F in islet transplant, we identified and validated a small molecule 5-HT 1F antagonist. This antagonist increased glucose stimulated insulin secretion from primary human islets and cAMP accumulation in primary human beta cells. Finally, the 5-HT 1F antagonist improved glycemia in marginal mass, human islet transplants into immunocompromised mice. We identify 5-HT 1F as a novel druggable target to improve human beta cell survival in the setting of islet transplantation.One Sentence Summary: Serotonin 1F receptor (5-HT 1F ) negatively regulates insulin secretion and beta cell survival during transplant.

    View details for DOI 10.1101/2024.05.01.591950

    View details for PubMedID 38746433

  • Vascular Ensheathment Reflects Characteristic Migratory Behavior of Paragangliomas. JCEM case reports Needleman, L., Holsinger, F. C., Annes, J. P. 2024; 2 (4): luae064

    View details for DOI 10.1210/jcemcr/luae064

    View details for PubMedID 38623531

    View details for PubMedCentralID PMC11017112

  • Caution on the Use of 68Ga-DOTATATE for the Diagnosis of Pheochromocytoma: A Report of 2 Cases. JCEM case reports Needleman, L., Enamandram, S., Annes, J. P. 2023; 1 (6): luad149


    Pheochromocytomas are intra-adrenal sympathetic neuroendocrine tumors that arise from chromaffin cells. Paragangliomas similarly arise from chromaffin cells, although at extra-adrenal sites such as sympathetic paraganglia in the abdomen/thorax, or parasympathetic paraganglia in the head/neck. Collectively, pheochromocytomas and paragangliomas are important to diagnose and resect because they may secrete harmful levels of catecholamines, have mass effects, hemorrhage, and/or metastasize. Anatomic imaging of pheochromocytomas is usually completed with computed tomography or magnetic resonance imaging; however, functional imaging may be used to provide additional localization, staging, and/or biologic information. Accordingly, selection of the proper functional imaging modality can be critical to developing the optimal therapeutic strategy. 68Gallium- and 64Copper-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-octreotate positron emission tomography computed tomography (68Ga- and 64Cu-DOTATATE) are widely used in evaluating pheochromocytomas and paragangliomas, although data regarding the sensitivity for diagnosing pheochromocytoma are limited. We report 2 cases of pheochromocytoma that showed nondiagnostic 68Ga-DOTATATE uptake but were subsequently visualized using alternative functional imaging modalities. Additionally, we provide a review of the literature to highlight the underappreciated limitations of functional adrenal imaging with somatostatin-based compounds.

    View details for DOI 10.1210/jcemcr/luad149

    View details for PubMedID 38045868

    View details for PubMedCentralID PMC10690848

  • Rare Presentation of Paroxysmal High B-Pee. Hypertension (Dallas, Tex. : 1979) Bradshaw, C., Abounasr, A., Brunsing, R. L., Kao, C. S., Reejhsinghani, R., Annes, J. P., Chung, B. I., Mihm, F., Bhalla, V. 2023

    View details for DOI 10.1161/HYPERTENSIONAHA.122.20790

    View details for PubMedID 36794582

  • On-demand electrochemically controlled compound release from an ultrasonically powered implant. RSC advances Wang, M. L., Chamberlayne, C. F., Xu, H., Mofidfar, M., Baltsavias, S., Annes, J. P., Zare, R. N., Arbabian, A. 2022; 12 (36): 23337-23345


    On-demand drug delivery systems are promising for a wide range of therapeutic applications. When combined with wireless implants for controlled drug delivery, they can reduce overall dosage and side effects. Here, we demonstrate release of fluorescein from a novel on-demand release system for negatively charged compounds. The release system is based on a modified electroresponsive polypyrrole nanoparticulate film designed to minimize ion exchange with the stored compound - a major passive leakage mechanism. We further designed an ultrasonically powered mm-sized implant to electronically control the on-demand drug delivery system in vivo. Release kinetics are characterized both in vitro and in vivo in mice using fluorescein as a model drug, demonstrating the feasibility of wireless, controllable drug release using an ultrasonically powered implant.

    View details for DOI 10.1039/d2ra03422k

    View details for PubMedID 36090393

    View details for PubMedCentralID PMC9382542

  • On-demand electrochemically controlled compound release from an ultrasonically powered implant RSC ADVANCES Wang, M. L., Chamberlayne, C. F., Xu, H., Mofidfar, M., Baltsavias, S., Annes, J. P., Zare, R. N., Arbabian, A. 2022; 12 (36): 23337-23345

    View details for DOI 10.1039/d2ra03422k

    View details for Web of Science ID 000841167500001

  • beta-Cell Succinate Dehydrogenase Deficiency Triggers Metabolic Dysfunction and Insulinopenic Diabetes. Diabetes Lee, S., Xu, H., Van Vleck, A., Mawla, A. M., Li, A. M., Ye, J., Huising, M. O., Annes, J. P. 2022


    Mitochondrial dysfunction plays a central role in Type 2 Diabetes (T2D); however, the pathogenic mechanisms in pancreatic beta-cells are incompletely elucidated. Succinate dehydrogenase (SDH) is a key mitochondrial enzyme with dual functions in the TCA cycle and electron transport chain (ETC). Using human diabetic samples and a mouse model of beta-cell-specific SDH ablation (SDHBbetaKO), we define SDH deficiency as a driver of mitochondrial dysfunction in beta-cell failure and insulinopenic diabetes. beta-Cell SDH deficiency impairs glucose-induced respiratory oxidative phosphorylation and mitochondrial membrane potential (DeltaPsim) collapse, thereby compromising glucose-stimulated ATP production, insulin secretion and beta-cell growth. Mechanistically, metabolomic and transcriptomic studies reveal that the loss of SDH causes excess succinate accumulation, which inappropriately activates mTORC1-regulated metabolic anabolism, including increased SREBP-regulated lipid synthesis. These alterations, which mirror diabetes-associated human beta-cell dysfunction, are partially reversed by acute mTOR inhibition with rapamycin. We propose SDH deficiency as a contributing mechanism to the progressive beta-cell failure of diabetes and identify mTORC1 inhibition as a potential mitigation strategy.

    View details for DOI 10.2337/db21-0834

    View details for PubMedID 35472723

  • SDHB knockout and succinate accumulation are insufficient for tumorigenesis but dual SDHB/NF1 loss yields SDHx-like pheochromocytomas. Cell reports Armstrong, N., Storey, C. M., Noll, S. E., Margulis, K., Soe, M. H., Xu, H., Yeh, B., Fishbein, L., Kebebew, E., Howitt, B. E., Zare, R. N., Sage, J., Annes, J. P. 2022; 38 (9): 110453


    Inherited pathogenic succinate dehydrogenase (SDHx) gene mutations cause the hereditary pheochromocytoma and paraganglioma tumor syndrome. Syndromic tumors exhibit elevated succinate, an oncometabolite that is proposed to drive tumorigenesis via DNA and histone hypermethylation, mitochondrial expansion, and pseudohypoxia-related gene expression. To interrogate this prevailing model, we disrupt mouse adrenal medulla SDHB expression, which recapitulates several key molecular features of human SDHx tumors, including succinate accumulation but not 5hmC loss, HIF accumulation, or tumorigenesis. By contrast, concomitant SDHB and the neurofibromin 1 tumor suppressor disruption yields SDHx-like pheochromocytomas. Unexpectedly, invivo depletion of the 2-oxoglutarate (2-OG) dioxygenase cofactor ascorbate reduces SDHB-deficient cell survival, indicating that SDHx loss may be better tolerated by tissues with high antioxidant capacity. Contrary to the prevailing oncometabolite model, succinate accumulation and 2-OG-dependent dioxygenase inhibition are insufficient for mouse pheochromocytoma tumorigenesis, which requires additional growth-regulatory pathway activation.

    View details for DOI 10.1016/j.celrep.2022.110453

    View details for PubMedID 35235785

  • Novel Pathogenic de novo INS p.T97P Variant Presenting with Severe Neonatal DKA. Endocrinology Lal, R. A., Moeller, H. P., Thomson, E. A., Horton, T. M., Lee, S., Freeman, R., Prahalad, P., Poon, A. S., Annes, J. P. 2021


    Pathogenic INS gene mutations are causative for Mutant INS-gene-induced Diabetes of Youth (MIDY). We characterize a novel de novo heterozygous INS gene mutation (c.289A>C, p.T97P) that presented in an autoantibody-negative 5-month-old male infant with severe diabetic ketoacidosis. In silico pathogenicity prediction tools provided contradictory interpretations, while structural modeling indicated a deleterious effect on proinsulin folding. Transfection of wildtype and INS p.T97P expression and luciferase reporter constructs demonstrated elevated intracellular mutant proinsulin levels and dramatically impaired proinsulin/insulin and luciferase secretion. Notably, proteasome inhibition partially and selectively rescued INS p.T97P-derived luciferase secretion. Additionally, expression of INS p.T97P caused increased intracellular proinsulin aggregate formation and XBP-1s protein levels, consistent with induction of endoplasmic reticulum stress. We conclude that INS p.T97P is a newly identified pathogenic A-chain variant that is causative for MIDY via disruption of proinsulin folding and processing with induction of the endoplasmic reticulum stress response.

    View details for DOI 10.1210/endocr/bqab246

    View details for PubMedID 34888628

  • Protocol for determining zinc-dependent beta cell-selective small-molecule delivery in mouse pancreas. STAR protocols Horton, T. M., Kraemer, B. R., Annes, J. P. 2021; 2 (1): 100263


    Targeted drug delivery to pancreatic islet beta cells is an unmet clinical need. beta cells possess a uniquely high Zn2+ concentration, and integrating Zn2+-binding activity into a small molecule can bias drug accumulation and activity toward beta cells. This protocol can be used to evaluate a molecule's capacity to chelate islet Zn2+, accumulate in islets, and stimulate beta cell-selective replication in mouse pancreas. One obstacle is establishing an LC-MS/MS-based method for compound measurement. Limitations include target compound ionizability and the time-sensitive nature of some experimental assay steps. For complete details on the use and execution of this protocol, please refer to Horton etal. (2019).

    View details for DOI 10.1016/j.xpro.2020.100263

    View details for PubMedID 33490979

  • Metastatic Paraganglioma of the Spine With SDHB Mutation: Case Report and Review of the Literature INTERNATIONAL JOURNAL OF SPINE SURGERY Jabarkheel, R., Pendharkar, A., Lavezo, J. L., Annes, J., Desai, K., Vogel, H., Desai, A. M. 2021; 14: S37–S45

    View details for DOI 10.14444/7163

    View details for Web of Science ID 000618077900008

  • Metastatic Paraganglioma of the Spine With SDHB Mutation: Case Report and Review of the Literature. International journal of spine surgery Jabarkheel, R., Pendharkar, A. V., Lavezo, J. L., Annes, J., Desai, K., Vogel, H., Desai, A. M. 2021; 14 (s4): S37-S45


    Paragangliomas (PGLs) are rare neuroendocrine tumors that can arise from any autonomic ganglion of the body. Most PGLs do not metastasize. Here, we present a rare case of metastatic PGL of the spine in a patient with a germline pathogenic succinate dehydrogenase subunit B (SDHB) mutation.In addition to a case report we provide a literature review of metastatic spinal PGL to highlight the importance of genetic testing and long-term surveillance of these patients.A 45-year-old woman with history of spinal nerve root PGL, 17 years prior, presented with back pain of several months' duration. Imaging revealed multilevel lytic lesions throughout the cervical, thoracic, and lumbar spine as well as involvement of the right mandibular condyle and clavicle. Percutaneous biopsy of the L1 spinal lesion confirmed metastatic PGL and the patient underwent posterior tumor resection and instrumented fusion of T7-T11. Postoperatively the patient was found to have a pathogenic SDHB deletion.Patients with SDHx mutation, particularly SDHB, have increased risk of developing metastatic PGLs. Consequently, these individuals require long-term surveillance given the risk for developing new tumors or disease recurrence, even years to decades after primary tumor resection. Surgical management of spinal metastatic PGL involves correcting spinal instability, minimizing tumor burden, and alleviating epidural cord compression. In patients with metastatic PGL of the spine, genetic testing should be considered.

    View details for DOI 10.14444/7163

    View details for PubMedID 33900943

  • A Wireless Implantable Potentiostat for Programmable Electrochemical Drug Delivery IEEE Biomedical Circuits and Systems (BIOCAS) Wang, M. L., Yeon, P., Chamberlayne, C. F., Mofidfar, M., Xu, H., Annes, J. P., Zare, R. N., Arbabian, A. 2021
  • Intracardiac paragangliomas: surgical approach and perioperative management. General thoracic and cardiovascular surgery Guenthart, B. A., Trope, W., Keeyapaj, W., Weiel, J. J., Edmonson, A., MacArthur, J. W., Annes, J. P., Woo, Y. J., Lui, N. S. 2020


    Intracardiac paragangliomas most commonly arise from the left atrium and are often infiltrative and densely adherent to surrounding structures. Given their rarity, only scattered reports exist in the literature and standardized perioperative and surgical management is not well established. We describe a case of a 60-year-old woman with a mildly functioning intracardiac paraganglioma in which division of the superior vena cava improved exposure and enabled a complex limited resection. Further, we provide an overview of the diagnostic workup, perioperative medical management, surgical approach, and surveillance strategy in patients with these challenging tumors.

    View details for DOI 10.1007/s11748-020-01503-2

    View details for PubMedID 33074472

  • Probability of positive genetic testing in patients diagnosed with pheochromocytoma and paraganglioma: Criteria beyond a family history. Surgery Alobuia, W. M., Ammar, S., Tyagi, M., Ghosh, C., Gunda, V., Annes, J. P., Kebebew, E. 2020


    BACKGROUND: Genetic testing for germline pheochromocytoma and paraganglioma susceptibility genes is associated with improved patient management. However, data are currently sparse on the probability of a positive testing result based on an individual's clinical presentation. This study evaluates clinical characteristics for association with testing positive for known pheochromocytoma and paraganglioma susceptibility genes.METHODS: This retrospective analysis examined 111 patients with a diagnosis of pheochromocytoma and paraganglioma who underwent genetic testing. Logistic regression and receiver operating characteristic analyses were performed to identify factors associated with a positive genetic testing result. Probabilities were then calculated for combinations of significant factors to determine the likelihood of a positive test result in each group.RESULTS: Of 32 patients with a family history of pheochromocytoma and paraganglioma, 31 (97%) had a germline mutation detected. Of 79 patients without a family history, 24 (30%) had a pathogenic germline mutation detected. In multivariate analysis, a positive family history, aged ≤47 years, and tumor size ≤2.9 cm were independent factors associated with a positive genetic testing result. Patients meeting all 3 criteria had a 100% probability compared with 13% in those without any of the criteria. In addition to a positive family history, having either aged ≤47 years or tumor size ≤2.9 cm resulted in a 90% and 100% probability of a positive result, respectively. In the absence of a family history, the probability in patients who were aged ≤47 years and had a tumor size ≤2.9 cm was 60%.CONCLUSION: In addition to a family history of pheochromocytoma and paraganglioma, aged ≤47 years, and tumor size ≤2.9 cm are associated with a higher probability of testing positive for a pheochromocytoma and paraganglioma susceptibility gene mutation. Patients meeting all 3 criteria have a 100% probability of a positive genetic testing result.

    View details for DOI 10.1016/j.surg.2020.08.027

    View details for PubMedID 33023754

  • Genetic testing in endocrine surgery: Opportunities for precision surgery. Surgery Alobuia, W., Annes, J., Kebebew, E. 2020


    Recent innovations in molecular and genetic diagnostic techniques have led to rapid advances in genomic medicine and their application to the clinic. The identification and classification of various genetic associations, syndromes, and susceptibility genes in endocrine surgical disorders are increasingly relevant to patient care. Hereditary endocrine disorders represent a significant proportion of disease encountered by endocrine surgeons. Hence, genetic testing has emerged as an important adjunct for the diagnosis and management of patients with endocrinesurgical disorders. This article summarizes commonly encountered inherited endocrine disorders and their tumor susceptibility genes, with a focus on the clinical utility of genetic testing and its impact on the surgical management of endocrine disorders.

    View details for DOI 10.1016/j.surg.2020.03.009

    View details for PubMedID 32376047

  • PAM staining intensity of primary neuroendocrine neoplasms is a potential prognostic biomarker. Scientific reports Horton, T. M., Sundaram, V. n., Lee, C. H., Hornbacker, K. n., Van Vleck, A. n., Benjamin, K. N., Zemek, A. n., Longacre, T. A., Kunz, P. L., Annes, J. P. 2020; 10 (1): 10943


    Neuroendocrine neoplasms (NENs) are rare epithelial tumors with heterogeneous and frequently unpredictable clinical behavior. Available biomarkers are insufficient to guide individual patient prognosis or therapy selection. Peptidylglycine α-amidating monooxygenase (PAM) is an enzyme expressed by neuroendocrine cells that participates in hormone maturation. The objective of this study was to assess the distribution, clinical associations and survival implications of PAM immunoreactivity in primary NENs. Of 109 primary NENs, 7% were PAM-negative, 25% were PAM-low and 68% were PAM-high. Staining intensity was high in small bowel (p = 0.04) and low in stomach (p = 0.004) NENs. PAM staining was lower in higher grade tumors (p < 0.001) and patients who died (p < 0.001) but did not vary by tumor size or stage at surgery. In patients who died, time to death was shorter in patients with reduced PAM immunoreactivity: median times to death were 11.3 (PAM-negative), 29.4 (PAM-low) and 61.7 (PAM-high) months. Lower PAM staining was associated with increased risk of death after adjusting for disease stage [PAM negative, HR = 13.8 (CI: 4.2-45.5)]. PAM immunoreactivity in primary NENs is readily assessable and a potentially useful stage-independent predictor of survival.

    View details for DOI 10.1038/s41598-020-68071-6

    View details for PubMedID 32616904

  • Generation of highly potent DYRK1A-dependent inducers of human beta-Cell replication via Multi-Dimensional compound optimization. Bioorganic & medicinal chemistry Allegretti, P. A., Horton, T. M., Abdolazimi, Y., Moeller, H. P., Yeh, B., Caffet, M., Michel, G., Smith, M., Annes, J. P. 2019: 115193


    Small molecule stimulation of beta-cell regeneration has emerged as a promising therapeutic strategy for diabetes. Although chemical inhibition of dual specificity tyrosine-phosphorylation-regulated kinase 1A (DYRK1A) is sufficient to enhance beta-cell replication, current lead compounds have inadequate cellular potency for in vivo application. Herein, we report the clinical stage anti-cancer kinase inhibitor OTS167 as a structurally novel, remarkably potent DYRK1A inhibitor and inducer of human beta-cell replication. Unfortunately, OTS167's target promiscuity and cytotoxicity curtails utility. To tailor kinase selectivity towards DYRK1A and reduce cytotoxicity we designed a library of fifty-one OTS167 derivatives based upon a modeled structure of the DYRK1A-OTS167 complex. Indeed, derivative characterization yielded several leads with exceptional DYRK1A inhibition and human beta-cell replication promoting potencies but substantially reduced cytotoxicity. These compounds are the most potent human beta-cell replication-promoting compounds yet described and exemplify the potential to purposefully leverage off-target activities of advanced stage compounds for a desired application.

    View details for DOI 10.1016/j.bmc.2019.115193

    View details for PubMedID 31757680

  • On-demand drug release from polypyrrole nanoparticulate films Chamberlayne, C., Baltsavias, S., Xu, H., Arbabian, A., Annes, J., Zare, R. AMER CHEMICAL SOC. 2019
  • Zinc-Chelating Small Molecules Preferentially Accumulate and Function within Pancreatic beta Cells CELL CHEMICAL BIOLOGY Horton, T. M., Allegretti, P. A., Lee, S., Moeller, H. P., Smith, M., Annes, J. P. 2019; 26 (2): 213-+
  • CC-401 Promotes beta-Cell Replication via Pleiotropic Consequences of DYRK1A/B Inhibition ENDOCRINOLOGY Abdolazimi, Y., Zhao, Z., Lee, S., Xu, H., Allegretti, P., Horton, T. M., Yeh, B., Moeller, H. P., Nichols, R. J., McCutcheon, D., Shalizi, A., Smith, M., Armstrong, N. A., Annes, J. P. 2018; 159 (9): 3143–57
  • CC-401 Promotes β-Cell Replication via Pleiotropic Consequences of DYRK1A/B Inhibition. Endocrinology Abdolazimi, Y. n., Lee, S. n., Xu, H. n., Allegretti, P. n., Horton, T. M., Yeh, B. n., Moeller, H. P., Nichols, R. J., McCutcheon, D. n., Shalizi, A. n., Smith, M. n., Armstrong, N. A., Annes, J. P. 2018


    Pharmacologic expansion of endogenous β-cells is a promising therapeutic strategy for diabetes. To elucidate the molecular pathways that control β-cell growth we screened ∼2,400 bioactive compounds for rat β-cell replication-modulating activity. Numerous hit compounds impaired or promoted rat β-cell replication, including CC-401, an advanced clinical candidate previously characterized as a c-Jun N-terminal kinase (JNK) inhibitor. Surprisingly, CC-401 induced rodent (in vitro and in vivo) and human (in vitro) β-cell replication via dual specificity tyrosine-phosphorylation-regulated kinases (DYRK1A/B) inhibition. In contrast to rat β-cells, which were broadly growth responsive to compound treatment, human β-cell replication was only consistently induced by DYRK1A/B inhibitors. This effect was enhanced by simultaneous glycogen synthase kinase-3β (GSK-3β) or transforming growth factor-β (ALK5/TGF-β) inhibition. Prior work emphasized DYRK1A/B inhibition-dependent activation of nuclear factor of activated T-cells (NFAT) as the primary mechanism of human β-cell replication induction. However, inhibition of NFAT activity had limited impact on CC-401-induced β-cell replication. Consequently, we investigated additional effects of CC-401-dependent DYRK1A/B inhibition. Indeed, CC-401 inhibited DYRK1A-dependent phosphorylation/stabilization of the β-cell replication-inhibitor p27Kip1. Additionally, CC-401 increased expression of numerous replication-promoting genes normally suppressed by the dimerization partner, RB-like, E2F and multi-vulval class B (DREAM) complex, which depends upon DYRK1A/B activity for integrity, including MYBL2 and FOXM1. In summary, we present a compendium of compounds as a valuable resource for manipulating the signaling pathways that control β-cell replication and leverage a novel DYRK1A/B inhibitor (CC-401) to expand our understanding of the molecular pathways that control β-cell growth.

    View details for PubMedID 29514186

  • Zinc-Chelating Small Molecules Preferentially Accumulate and Function within Pancreatic β Cells. Cell chemical biology Horton, T. M., Allegretti, P. A., Lee, S. n., Moeller, H. P., Smith, M. n., Annes, J. P. 2018


    Diabetes is a hyperglycemic condition characterized by pancreatic β-cell dysfunction and depletion. Whereas methods for monitoring β-cell function in vivo exist, methods to deliver therapeutics to β cells are lacking. We leveraged the rare ability of β cells to concentrate zinc to preferentially trap zinc-binding molecules within β cells, resulting in β-cell-targeted compound delivery. We determined that zinc-rich β cells and islets preferentially accumulated TSQ (6-methoxy-8-p-toluenesulfonamido-quinoline) in a zinc-dependent manner compared with exocrine pancreas. Next, we asked whether appending a zinc-chelating moiety onto a β-cell replication-inducing compound was sufficient to confer preferential β-cell accumulation and activity. Indeed, the hybrid compound preferentially accumulated within rodent and human islets in a zinc-dependent manner and increased the selectivity of replication-promoting activity toward β cells. These data resolve the fundamental question of whether intracellular accumulation of zinc-chelating compounds is influenced by zinc content. Furthermore, application of this principle yielded a proof-of-concept method for β-cell-targeted drug delivery and bioactivity.

    View details for PubMedID 30527998

  • Genetic Disruption of Adenosine Kinase in Mouse Pancreatic ß-Cells Protects Against High Fat Diet-Induced Glucose Intolerance. Diabetes Navarro, G., Abdolazami, Y., Zhao, Z., Xu, H., Lee, S., Armstrong, N. A., Annes, J. P. 2017


    Islet β-cells adapt to insulin resistance through increased insulin secretion and expansion. Type 2 diabetes typically occurs when prolonged insulin resistance exceeds the adaptive capacity of β-cells. Our prior screening efforts led to the discovery that adenosine kinase (ADK) inhibitors stimulate β-cell replication. Here, we evaluated whether ADK disruption in mouse β-cells affects β-cell mass and/or protects against high-fat diet (HFD)-induced glucose dysregulation. Mice targeted at the Adk locus were bred to Rip-Cre and Ins1-Cre/ERT(1Lphi) mice to enable constitutive (βADKO) and conditional (iβADKO) disruption of ADK expression in β-cells, respectively. Weight gain, glucose tolerance, insulin sensitivity, and glucose-stimulated insulin secretion (GSIS) were longitudinally monitored in normal chow (NC)-fed and HFD-fed mice. In addition, β-cell mass and replication were measured by immunofluorescence-based islet morphometry. NC-fed adult βADKO and iβADKO mice displayed glucose tolerance, insulin tolerance and β-cell mass comparable to control animals. By contrast, HFD-fed βADKO and iβADKO animals had improved glucose tolerance and increased in vivo GSIS. Improved glucose handling was associated with increased β-cell replication and mass. We conclude that ADK expression negatively regulates the adaptive β-cell response to HFD challenge. Therefore, modulation of ADK activity is a potential strategy for enhancing the adaptive β-cell response.

    View details for DOI 10.2337/db16-0816

    View details for PubMedID 28468960

  • Hyaluronan content governs tissue stiffness in pancreatic islet inflammation. The Journal of biological chemistry Nagy, N. n., de la Zerda, A. n., Kaber, G. n., Johnson, P. Y., Hu, K. H., Kratochvil, M. J., Yadava, K. n., Zhao, W. n., Cui, Y. n., Navarro, G. n., Annes, J. P., Wight, T. N., Heilshorn, S. C., Bollyky, P. L., Butte, M. J. 2017


    We have identified a novel role for hyaluronan (HA), an extracellular matrix (ECM) polymer, in governing the mechanical properties of inflamed tissues. We recently reported that insulitis in type 1 diabetes (T1D) of mice and humans is preceded by intra-islet accumulation of HA, a highly hygroscopic polymer. Using the DORmO double transgenic (DO11.10 x RIPmOVA) mouse model of T1D, we asked whether autoimmune insulitis was associated with changes in the stiffness of islets. To measure islet stiffness, we used atomic force microscopy (AFM) and developed a novel "bed of nails"-like approach that uses quartz glass nanopillars to anchor islets, solving a long-standing problem of keeping tissue-scale objects immobilized while performing AFM. We measured stiffness via AFM nanoindentation with a spherical indenter and found that insulitis made islets mechanically soft compared to controls. Conversely, treatment with 4-methylumbelliferone (4-MU), a small-molecule inhibitor of HA synthesis, reduced HA accumulation, diminished swelling, and restored basal tissue stiffness. These results indicate that HA content governs the mechanical properties of islets. In hydrogels with variable HA content we confirmed that increased HA leads to mechanically softer hydrogels, consistent with our model. In light of recent reports that the insulin production of islets is mechanosensitive, these findings open up an exciting new avenue of research into the fundamental mechanisms by which inflammation impacts local cellular responses.

    View details for PubMedID 29183997

  • Electrically controlled release of insulin using polypyrrole nanoparticles NANOSCALE Hosseini-Nassab, N., Samanta, D., Abdolazimi, Y., Annes, J. P., Zare, R. N. 2017; 9 (1): 143-149


    Conducting polymers present an opportunity for developing programmable, adjustable, spatially, and temporally controllable drug delivery systems. While several small molecule drugs have been released from thin conductive polymeric films successfully, delivering large molecule therapeutics, such as polypeptides and nucleic acids, has remained a significant challenge. Poor drug loading (∼ng cm(-2)) of thin films coupled with film instability has, in many cases, made conducting polymer films refractory to clinical development. To address these limitations, we have utilized conductive polymer nanoparticulate backbones to controllably release insulin, a high molecular weight, clinically relevant polypeptide. We find that the interaction between insulin and the polymer scaffold can be described by a simple Langmuir-type adsorption model. By modifying the ratio of the amount of nanoparticles to the amount of insulin, we have obtained drug loading percentages estimated to be as high as 51 wt% percent. In vivo experiments in mice confirmed retained bioactivity of the released insulin after electrical stimulation.

    View details for DOI 10.1039/c6nr08288b

    View details for Web of Science ID 000391739300020

    View details for PubMedID 27929180

    View details for PubMedCentralID PMC5215613

  • A High-content In Vitro Pancreatic Islet ß-cell Replication Discovery Platform. Journal of visualized experiments : JoVE Zhao, Z., Abdolazimi, Y., Armstrong, N. A., Annes, J. P. 2016


    Loss of insulin-producing β-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion of endogenous insulin-producing pancreatic islet β-cells remains an attractive strategy. β-cells have limited spontaneous regenerative activity; consequently, a crucial research effort is to develop a precise understanding of the molecular pathways that restrain β-cell growth and to identify drugs capable of overcoming these restraints. Herein an automated high-content image-based primary-cell screening method to identify β-cell replication-promoting small molecules is presented. Several, limitations of prior methodologies are surmounted. First, use of primary islet cells rather than an immortalized cell-line maximizes retention of in vivo growth restraints. Second, use of mixed-composition islet-cell cultures rather than a β-cell-line allows identification of both lineage-restricted and general growth stimulators. Third, the technique makes practical the use of primary islets, a limiting resource, through use of a 384-well format. Fourth, detrimental experimental variability associated with erratic islet culture quality is overcome through optimization of isolation, dispersion, plating and culture parameters. Fifth, the difficulties of accurately and consistently measuring the low basal replication rate of islet endocrine-cells are surmounted with optimized immunostaining parameters, automated data acquisition and data analysis; automation simultaneously enhances throughput and limits experimenter bias. Notable limitations of this assay are the use of dispersed islet cultures which disrupts islet architecture, the use of rodent rather than human islets and the inherent limitations of throughput and cost associated with the use of primary cells. Importantly, the strategy is easily adapted for human islet replication studies. This assay is well suited for investigating the mitogenic effect of substances on β-cells and the molecular mechanisms that regulate β-cell growth.

    View details for DOI 10.3791/54298

    View details for PubMedID 27500720

  • Repurposing cAMP-Modulating Medications to Promote beta-Cell Replication MOLECULAR ENDOCRINOLOGY Zhao, Z., Low, Y. S., Armstrong, N. A., Ryu, J. H., Sun, S. A., Arvanites, A. C., Hollister-Lock, J., Shah, N. H., Weir, G. C., Annes, J. P. 2014; 28 (10): 1682-1697


    Loss of β-cell mass is a cardinal feature of diabetes. Consequently, developing medications to promote β-cell regeneration is a priority. 3'-5'-Cyclic adenosine monophosphate (cAMP) is an intracellular second messenger that modulates β-cell replication. We investigated whether medications that increase cAMP stability or synthesis selectively stimulate β-cell growth. To identify cAMP stabilizing medications that promote β-cell replication we performed high-content screening of a phosphodiesterase-inhibitor (PDE-I) library. PDE3,4 and 10 inhibitors, including dipyridamole, were found to promote β-cell replication in an adenosine receptor-dependent manner. Dipyridamole's action is specific for β-cells and not α-cells. Next we demonstrated that norepinephrine (NE), a physiologic suppressor of cAMP synthesis in β-cells, impairs β-cell replication via activation of α2-adrenergic receptors. Accordingly, mirtazapine, an α2-adrenergic receptor antagonist and antidepressant, prevents NE-dependent suppression of β-cell replication. Interestingly, NE's growth-suppressive effect is modulated by endogenously expressed catecholamine-inactivating enzymes (COMT and MAO) and is dominant over the growth-promoting effects of PDE-Is. Treatment with dipyridamole and/or mirtazapine promote β-cell replication in mice and treatment with dipyridamole is associated with reduced glucose levels in humans. This work provides new mechanistic insights into cAMP-dependent growth regulation of β-cells and highlights the potential of commonly prescribed medications to influence β-cell growth.

    View details for DOI 10.1210/me.2014-1120

    View details for Web of Science ID 000346837000010

    View details for PubMedID 25083741

  • Adult tissue sources for new beta cells TRANSLATIONAL RESEARCH Nichols, R. J., New, C., Annes, J. P. 2014; 163 (4): 418-431


    The diabetes pandemic incurs extraordinary public health and financial costs that are projected to expand for the foreseeable future. Consequently, the development of definitive therapies for diabetes is a priority. Currently, a wide spectrum of therapeutic strategies-from implantable insulin delivery devices to transplantation-based cell replacement therapy, to β-cell regeneration-focus on replacing the lost insulin-producing capacity of individuals with diabetes. Among these, β-cell regeneration remains promising but heretofore unproved. Indeed, recent experimental work has uncovered surprising biology that underscores the potential therapeutic benefit of β-cell regeneration. These studies have elucidated a variety of sources for the endogenous production of new β cells from existing cells. First, β cells, long thought to be postmitotic, have demonstrated the potential for regenerative capacity. Second, the presence of pancreatic facultative endocrine progenitor cells has been established. Third, the malleability of cellular identity has availed the possibility of generating β cells from other differentiated cell types. Here, we review the exciting developments surrounding endogenous sources of β-cell production and consider the potential of realizing a regenerative therapy for diabetes from adult tissues.

    View details for DOI 10.1016/j.trsl.2013.11.012

    View details for Web of Science ID 000334648500015

    View details for PubMedID 24345765

  • A liver Hif-2a-Irs2 pathway sensitizes hepatic insulin signaling and is modulated by Vegf inhibition. Nature medicine Wei, K., Piecewicz, S. M., McGinnis, L. M., Taniguchi, C. M., Wiegand, S. J., Anderson, K., Chan, C. W., Mulligan, K. X., Kuo, D., Yuan, J., Vallon, M., Morton, L. C., Lefai, E., Simon, M. C., Maher, J. J., Mithieux, G., Rajas, F., Annes, J. P., McGuinness, O. P., Thurston, G., Giaccia, A. J., Kuo, C. J. 2013; 19 (10): 1331-1337


    Insulin initiates diverse hepatic metabolic responses, including gluconeogenic suppression and induction of glycogen synthesis and lipogenesis. The liver possesses a rich sinusoidal capillary network with a higher degree of hypoxia and lower gluconeogenesis in the perivenous zone as compared to the rest of the organ. Here, we show that diverse vascular endothelial growth factor (VEGF) inhibitors improved glucose tolerance in nondiabetic C57BL/6 and diabetic db/db mice, potentiating hepatic insulin signaling with lower gluconeogenic gene expression, higher glycogen storage and suppressed hepatic glucose production. VEGF inhibition induced hepatic hypoxia through sinusoidal vascular regression and sensitized liver insulin signaling through hypoxia-inducible factor-2α (Hif-2α, encoded by Epas1) stabilization. Notably, liver-specific constitutive activation of HIF-2α, but not HIF-1α, was sufficient to augment hepatic insulin signaling through direct and indirect induction of insulin receptor substrate-2 (Irs2), an essential insulin receptor adaptor protein. Further, liver Irs2 was both necessary and sufficient to mediate Hif-2α and Vegf inhibition effects on glucose tolerance and hepatic insulin signaling. These results demonstrate an unsuspected intersection between Hif-2α-mediated hypoxic signaling and hepatic insulin action through Irs2 induction, which can be co-opted by Vegf inhibitors to modulate glucose metabolism. These studies also indicate distinct roles in hepatic metabolism for Hif-1α, which promotes glycolysis, and Hif-2α, which suppresses gluconeogenesis, and suggest new treatment approaches for type 2 diabetes mellitus.

    View details for DOI 10.1038/nm.3295

    View details for PubMedID 24037094

  • A liver Hif-2 alpha-Irs2 pathway sensitizes hepatic insulin signaling and is modulated by Vegf inhibition NATURE MEDICINE Wei, K., Piecewicz, S. M., McGinnis, L. M., Taniguchi, C. M., Wiegand, S. J., Anderson, K., Chan, C. W., Mulligan, K. X., Kuo, D., Yuan, J., Vallon, M., Morton, L. C., Lefai, E., Simon, M. C., Maher, J. J., Mithieux, G., Rajas, F., Annes, J. P., McGuinness, O. P., Thurston, G., Giaccia, A. J., Kuo, C. J. 2013; 19 (10): 1331-?


    Insulin initiates diverse hepatic metabolic responses, including gluconeogenic suppression and induction of glycogen synthesis and lipogenesis. The liver possesses a rich sinusoidal capillary network with a higher degree of hypoxia and lower gluconeogenesis in the perivenous zone as compared to the rest of the organ. Here, we show that diverse vascular endothelial growth factor (VEGF) inhibitors improved glucose tolerance in nondiabetic C57BL/6 and diabetic db/db mice, potentiating hepatic insulin signaling with lower gluconeogenic gene expression, higher glycogen storage and suppressed hepatic glucose production. VEGF inhibition induced hepatic hypoxia through sinusoidal vascular regression and sensitized liver insulin signaling through hypoxia-inducible factor-2α (Hif-2α, encoded by Epas1) stabilization. Notably, liver-specific constitutive activation of HIF-2α, but not HIF-1α, was sufficient to augment hepatic insulin signaling through direct and indirect induction of insulin receptor substrate-2 (Irs2), an essential insulin receptor adaptor protein. Further, liver Irs2 was both necessary and sufficient to mediate Hif-2α and Vegf inhibition effects on glucose tolerance and hepatic insulin signaling. These results demonstrate an unsuspected intersection between Hif-2α-mediated hypoxic signaling and hepatic insulin action through Irs2 induction, which can be co-opted by Vegf inhibitors to modulate glucose metabolism. These studies also indicate distinct roles in hepatic metabolism for Hif-1α, which promotes glycolysis, and Hif-2α, which suppresses gluconeogenesis, and suggest new treatment approaches for type 2 diabetes mellitus.

    View details for DOI 10.1038/nm.3295

    View details for Web of Science ID 000325531700034

    View details for PubMedID 24037094

    View details for PubMedCentralID PMC3795838

  • The influence of sodium- and calcium-regulatory hormone interventions on adipocytokines in obesity and diabetes. Metabolism Vaidya, A., Underwood, P. C., Annes, J. P., Sun, B., Williams, G. H., Forman, J. P., Williams, J. S. 2013; 62 (4): 539-547


    The renin-angiotensin-aldosterone system (RAAS), vitamin D, and parathyroid hormone have all been implicated as regulators of adipocytokines and inflammation. We evaluated human interventional study protocols to investigate whether controlled modulations of these calcium- and sodium-regulatory hormones could influence adipocytokines and inflammation in obesity and diabetes.Post-hoc analyses of two separate human protocols (Protocol 1, n=14; Protocol 2, n=24) conducted in a clinical research setting after rigorous control of diet, posture, medications, and diurnal rhythm, were performed. Protocol 1 evaluated obese hypertensives with vitamin D deficiency who received an infusion of angiotensin II (AngII) before and after 1month of vitamin D3 therapy. Protocol 2 evaluated obese subjects with type 2 diabetes who also received AngII. Adipocytokines and inflammatory markers were measured before and after vitamin D3 therapy, and also before and after infusions of AngII.Vitamin D3 therapy significantly raised 25(OH)D and 1,25(OH)2D concentrations, and lowered parathyroid hormone, but had no effect on concentrations of adiponectin, resistin, leptin, IL-6, PAI-1, urinary TGFβ1, or HOMA-IR. AngII infusions, despite significant elevations in blood pressure and serum aldosterone, did not influence adipocytokine concentrations in either protocol.In contrast to prior studies conducted in healthy populations, or those that could not control major regulators of the RAAS or adipocytokines, we observed that robust modulations in calcium- and sodium-regulatory hormones did not influence adipocytokines or inflammation in obesity or diabetes. Adipose-tissue physiology in these conditions may alter the hormonal regulation of inflammatory parameters.

    View details for DOI 10.1016/j.metabol.2012.10.001

    View details for PubMedID 23142162

    View details for PubMedCentralID PMC3572332

  • In Vivo Screening for Secreted Proteins That Modulate Glucose Handling Identifies Interleukin-6 Family Members as Potent Hypoglycemic Agents PLOS ONE Chen, C. A., Carolan, P. C., Annes, J. P. 2012; 7 (9)


    Diabetes is a disease of abnormal glucose homeostasis characterized by chronic hyperglycemia and a broad array of consequent organ damage. Because normal glucose homeostasis is maintained by a complex interaction between behavior (feeding and physical activity) and metabolic activity that is modulated by inter-organ signaling through secreted factors, disease modeling in vitro is necessarily limited. In contrast, in vivo studies allow complex metabolic phenotypes to be studied but present a barrier to high throughput studies. Here we present the development of a novel in vivo screening platform that addresses this primary limitation of in vivo experimentation. Our platform leverages the large secretory capacity of the liver and the hepatocyte transfection technique of hydrodynamic tail vein injection to achieve supraphysiologic blood levels of secreted proteins. To date, the utility of hydrodynamic transfection has been limited by the deleterious impact of the variable transfection efficiency inherent to this technique. We overcome this constraint by co-transfection of a secreted luciferase cDNA whose product can be easily monitored in the blood of a living animal and used as a surrogate marker for transfection efficiency and gene expression levels. To demonstrate the utility of our strategy, we screened 248 secreted proteins for the ability to enhance glucose tolerance. Surprisingly, interleukin-6 and several of its family members but not other well-recognized insulin sensitizing agents were identified as potent hypoglycemic factors. We propose this experimental system as a powerful and flexible in vivo screening platform for identifying genes that modulate complex behavioral and metabolic phenotypes.

    View details for DOI 10.1371/journal.pone.0044600

    View details for Web of Science ID 000308577600073

    View details for PubMedID 22962620

    View details for PubMedCentralID PMC3433445

  • Genetics of adrenocortical disease: an update CURRENT OPINION IN ENDOCRINOLOGY DIABETES AND OBESITY Bar-Lev, A., Annes, J. P. 2012; 19 (3): 159-167


    Disease states characterized by abnormal cellular function or proliferation frequently reflect aberrant genetic information. By revealing disease-specific DNA mutations, we gain insight into normal physiology, pathophysiology, potential therapeutic targets and are better equipped to evaluate an individual's disease risks. This review examines recent advances in our understanding of the genetic basis of adrenal cortical disease.Important advances made in the past year have included identification of KCNJ5 potassium channel mutations in the pathogenesis of both aldosterone-producing adenomas and familial hyperaldosteronism type III; characterization of phosphodiesterase 11A as a modifier of phenotype in Carney complex caused by protein kinase, cAMP-dependent, regulatory subunit, type-I mutations; the finding of 11β-hydroxysteroid dehydrogenase type I mutations as a novel mechanism for cortisone reductase deficiency; and demonstration of potential mortality benefit in pursuing comprehensive presymptomatic screening for patients with Li-Fraumeni syndrome, including possible reduction in risks associated with adrenocortical carcinoma.This research review provides a framework for the endocrinologist to maintain an up-to-date understanding of adrenal cortical disease genetics.

    View details for DOI 10.1097/MED.0b013e328352f013

    View details for Web of Science ID 000303255900003

    View details for PubMedID 22476103

  • Adenosine kinase inhibition selectively promotes rodent and porcine islet beta-cell replication PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Annes, J. P., Ryu, J. H., Lam, K., Carolan, P. J., Utz, K., Hollister-Lock, J., Arvanites, A. C., Rubin, L. L., Weir, G., Melton, D. A. 2012; 109 (10): 3915-3920


    Diabetes is a pathological condition characterized by relative insulin deficiency, persistent hyperglycemia, and, consequently, diffuse micro- and macrovascular disease. One therapeutic strategy is to amplify insulin-secretion capacity by increasing the number of the insulin-producing β cells without triggering a generalized proliferative response. Here, we present the development of a small-molecule screening platform for the identification of molecules that increase β-cell replication. Using this platform, we identify a class of compounds [adenosine kinase inhibitors (ADK-Is)] that promote replication of primary β cells in three species (mouse, rat, and pig). Furthermore, the replication effect of ADK-Is is cell type-selective: treatment of islet cell cultures with ADK-Is increases replication of β cells but not that of α cells, PP cells, or fibroblasts. Short-term in vivo treatment with an ADK-I also increases β-cell replication but not exocrine cell or hepatocyte replication. Therefore, we propose ADK inhibition as a strategy for the treatment of diabetes.

    View details for DOI 10.1073/pnas.1201149109

    View details for Web of Science ID 000301117700063

    View details for PubMedID 22345561

    View details for PubMedCentralID PMC3309788

  • Erdheim-Chester disease presenting with cutaneous involvement: a case report and literature review JOURNAL OF CUTANEOUS PATHOLOGY Volpicelli, E. R., Doyle, L., Annes, J. P., Murray, M. F., Jacobsen, E., Murphy, G. F., Saavedra, A. P. 2011; 38 (3): 280-285


    Erdheim-Chester disease (ECD) is a rare, systemic, non-familial histiocytic disorder, first described by Jakob Erdheim and William Chester in 1930. Most patients have multiple sites of involvement at presentation. The most common site of involvement is the long bones of the axial skeleton, which is seen almost universally, followed by the nervous system, heart, lungs, orbit and retroperitoneum, which are seen in up to 50% of cases. Cutaneous involvement is rarely a presenting symptom of ECD, with two reported cases in the English literature. The diagnosis of ECD is rarely made by skin biopsy because of the relative rarity of cutaneous involvement as a presenting feature, and also perhaps because of the difficulty in distinguishing the histopathological appearance from potential mimics. The importance of distinguishing ECD from other cutaneous disorders with similar pathology lies in the implications for both treatment and prognosis. ECD is an aggressive, often fatal disorder, with death from disease occurring in greater than 50% of patients.

    View details for DOI 10.1111/j.1600-0560.2010.01650.x

    View details for Web of Science ID 000286380300006

    View details for PubMedID 21143617

    View details for PubMedCentralID PMC3619727

  • Risks of Presymptomatic Direct-to-Consumer Genetic Testing. NEW ENGLAND JOURNAL OF MEDICINE Annes, J. P., Giovanni, M. A., Murray, M. F. 2010; 363 (12): 1100-1101

    View details for DOI 10.1056/NEJMp1006029

    View details for Web of Science ID 000281795800003

    View details for PubMedID 20843242

  • Integrin alpha(v)beta(6)-mediated activation of latent TGF-beta requires the latent TGF-beta binding protein-1 JOURNAL OF CELL BIOLOGY Annes, J. P., Chen, Y., Munger, J. S., Rifkin, D. B. 2004; 165 (5): 723-734


    Transforming growth factor-betas (TGF-beta) are secreted as inactive complexes containing the TGF-beta, the TGF-beta propeptide, also called the latency-associated protein (LAP), and the latent TGF-beta binding protein (LTBP). Extracellular activation of this complex is a critical but incompletely understood step in TGF-beta regulation. We have investigated the role of LTBP in modulating TGF-beta generation by the integrin alphaVbeta6. We show that even though alphavbeta6 recognizes an RGD on LAP, LTBP-1 is required for alphaVbeta6-mediated latent TGF-beta activation. The domains of LTBP-1 necessary for activation include the TGF-beta propeptide-binding domain and a basic amino acid sequence (hinge domain) with ECM targeting properties. Our results demonstrate an LTBP-1 isoform-specific function in alphaVbeta6-mediated latent TGF-beta activation; LTBP-3 is unable to substitute for LTBP-1 in this assay. The results reveal a functional role for LTBP-1 in latent TGF-beta activation and suggest that activation of specific latent complexes is regulated by distinct mechanisms that may be determined by the LTBP isoform and its potential interaction with the matrix.

    View details for DOI 10.1083/jcb.200312172

    View details for Web of Science ID 000221986200013

    View details for PubMedID 15184403

    View details for PubMedCentralID PMC2172370

  • A genetic screen to identify latent transforming growth factor beta activators ANALYTICAL BIOCHEMISTRY Annes, J., Vassallo, M., Munger, J. S., Rifkin, D. B. 2004; 327 (1): 45-54


    The mechanisms by which latent transforming growth factor beta (TGFbeta) is converted to the active cytokine are largely unknown. Here we present a genetic screen that combines retroviral mutagenesis and cDNA expression cloning to reveal proteins involved in the extracellular regulation of latent TGFbeta activation. The screen employs a cell line engineered to express green fluorescent protein (GFP) in response to TGFbeta. The cells produce their own latent TGFbeta. Therefore, after transduction with a retroviral cDNA library that contains an insert for an activator of latent TGFbeta, cells expressing the activator are GFP-bright. These cells are enriched by fluorescence-activated cell sorting and grown as individual clones. The isolated clones are cocultured with a second TGFbeta reporter cell line that produces luciferase in response to TGFbeta. Cells that have acquired the ability to activate latent TGFbeta induce luciferase expression in the absence but not in the presence of neutralizing antibodies to TGFbeta. The activator expressed by the positive clones can be identified by retrieval of the retrovirus cDNA insert.

    View details for DOI 10.1016/j.ab.2003.11.029

    View details for Web of Science ID 000220535400005

    View details for PubMedID 15033509

  • Annexin II-mediated plasmin generation activates TGF-beta 3 during epithelial-mesenchymal transformation in the developing avian heart DEVELOPMENTAL BIOLOGY Krishnan, S., Deora, A. B., Annes, J. P., Osoria, J., RIFKIN, D. B., Hajar, K. A. 2004; 265 (1): 140-154


    Epithelial-mesenchymal transformation (EMT), the process by which epithelial cells are converted into motile, invasive mesenchymal cells, is critical to valvulogenesis. Transforming growth factor-beta3 (TGF-beta3), an established mediator of avian atrioventricular (AV) canal EMT, is secreted as a latent complex. In vitro, plasmin-mediated proteolysis has been shown to release active TGF-betas from the latent complex. Annexin II, a co-receptor for tissue plasminogen activator (tPA) and plasminogen, promotes cell-surface generation of the serine protease plasmin. Here, we show that annexin II-mediated plasmin activity regulates release of active TGF-beta3 during chick AV canal EMT. Primary embryonic endocardial-derived cells express annexin II which promotes plasminogen activation in vitro. Incubation of heart explant cultures with either alpha(2)antiplasmin (alpha(2)AP), a major physiological plasmin inhibitor, or anti-annexin II IgG, blocked EMT by approximately 80%, and 50%, respectively. Anti-annexin II IgG-mediated inhibition of EMT was overcome by the addition of recombinant TGF-beta3. Upon treatment with anti-annexin II IgG or alpha(2)AP, conditioned medium from heart explant cultures showed absence of the active fragment of TGF-beta3 by Western blot analysis and a approximately 50% decrease in TGF-beta specific bioactivity. Our results suggest that annexin II-mediated plasmin activity regulates the release of active TGF-beta during cardiac valve development in the avian heart.

    View details for DOI 10.1016/j.ydbio.2003.08.026

    View details for Web of Science ID 000187669700012

    View details for PubMedID 14697359

  • Making sense of latent TGF beta activation JOURNAL OF CELL SCIENCE Annes, J. P., Munger, J. S., RIFKIN, D. B. 2003; 116 (2): 217-224


    TGFbeta is secreted as part of a latent complex that is targeted to the extracellular matrix. A variety of molecules, 'TGFbeta activators,' release TGFbeta from its latent state. The unusual temporal discontinuity of TGFbeta synthesis and action and the panoply of TGFbeta effects contribute to the interest in TGF-beta. However, the logical connections between TGFbeta synthesis, storage and action are obscure. We consider the latent TGFbeta complex as an extracellular sensor in which the TGFbeta propeptide functions as the detector, latent-TGFbeta-binding protein (LTBP) functions as the localizer, and TGF-beta functions as the effector. Such a view provides a logical continuity for various aspects of TGFbeta biology and allows us to appreciate TGFbeta biology from a new perspective.

    View details for DOI 10.1242/jcs.00229

    View details for Web of Science ID 000180677200002

    View details for PubMedID 12482908

  • Latent TGF-beta binding protein-3 (LTBP-3) requires binding to TGF-beta for secretion FEBS LETTERS Chen, Y., Dabovic, B., Annes, J. P., Rifkin, D. B. 2002; 517 (1-3): 277-280


    Latent transforming growth factor-beta (TGF-beta) binding protein (LTBP)-1, which is easily secreted, has been shown to enhance the secretion of TGF-beta. Here we show that another member of the LTBP family, LTBP-3, is not secreted by several cell types, but secretion occurs after coexpression with TGF-beta. The secretion of LTBP-3 requires complexing of LTBP-3 with Cys33 of the TGF-beta propeptide.

    View details for Web of Science ID 000175225600055

    View details for PubMedID 12062452

  • The integrin alpha(V)beta(6) binds and activates latent TGF beta 3 FEBS LETTERS Annes, J. P., RIFKIN, D. B., Munger, J. S. 2002; 511 (1-3): 65-68


    Transforming growth factors-beta (TGFbeta1, 2 and 3) are secreted in a complex with their propeptides (latency-associated peptide 1 (LAP1), 2 and 3). TGFbeta signaling requires the dissociation of LAP and TGFbeta, a process termed latent TGFbeta activation. This process is a critical but incompletely understood step in the regulation of TGFbeta function. In particular, the extent to which activation mechanisms differ among the three TGFbeta isoforms is relatively unexplored. We show here that alphaVbeta6 binds and activates latent TGFbeta3.

    View details for Web of Science ID 000173628000013

    View details for PubMedID 11821050

  • The latent transforming growth factor-beta-binding protein-1 promotes in vitro differentiation of embryonic stem cells into endothelium MOLECULAR BIOLOGY OF THE CELL Gualandris, A., Annes, J. P., Arese, M., Noguera, I., Jurukovski, V., Rifkin, D. B. 2000; 11 (12): 4295-4308


    The latent transforming growth factor-beta-binding protein-1 (LTBP-1) belongs to a family of extracellular glycoproteins that includes three additional isoforms (LTBP-2, -3, and -4) and the matrix proteins fibrillin-1 and -2. Originally described as a TGF-beta-masking protein, LTBP-1 is involved both in the sequestration of latent TGF-beta in the extracellular matrix and the regulation of its activation in the extracellular environment. Whereas the expression of LTBP-1 has been analyzed in normal and malignant cells and rodent and human tissues, little is known about LTBP-1 in embryonic development. To address this question, we used murine embryonic stem (ES) cells to analyze the appearance and role of LTBP-1 during ES cell differentiation. In vitro, ES cells aggregate to form embryoid bodies (EBs), which differentiate into multiple cell lineages. We analyzed LTBP-1 gene expression and LTBP-1 fiber appearance with respect to the emergence and distribution of cell types in differentiating EBs. LTBP-1 expression increased during the first 12 d in culture, appeared to remain constant between d 12 and 24, and declined thereafter. By immunostaining, fibrillar LTBP-1 was observed in those regions of the culture containing endothelial, smooth muscle, and epithelial cells. We found that inclusion of a polyclonal antibody to LTBP-1 during EB differentiation suppressed the expression of the endothelial specific genes ICAM-2 and von Willebrand factor and delayed the organization of differentiated endothelial cells into cord-like structures within the growing EBs. The same effect was observed when cultures were treated with either antibodies to TGF-beta or the latency associated peptide, which neutralize TGF-beta. Conversely, the organization of endothelial cells was enhanced by incubation with TGF-beta 1. These results suggest that during differentiation of ES cells LTBP-1 facilitates endothelial cell organization via a TGF-beta-dependent mechanism.

    View details for Web of Science ID 000165955000018

    View details for PubMedID 11102524

    View details for PubMedCentralID PMC15073

  • PKC-theta is required for TCR-induced NF-kappa B activation in mature but not immature T lymphocytes NATURE Sun, Z. M., Arendt, C. W., Ellmeier, W., Schaeffer, E. M., Sunshine, M. J., Gandhi, L., Annes, J., Petrzilka, D., Kupfer, A., Schwartzberg, P. L., Littman, D. R. 2000; 404 (6776): 402-407


    Productive interaction of a T lymphocyte with an antigen-presenting cell results in the clustering of the T-cell antigen receptor (TCR) and the recruitment of a large signalling complex to the site of cell-cell contact. Subsequent signal transduction resulting in cytokine gene expression requires the activation of one or more of the multiple isoenzymes of serine/threonine-specific protein kinase C (PKC). Among the several PKC isoenzymes expressed in T cells, PKC-theta is unique in being rapidly recruited to the site of TCR clustering. Here we show that PKC-theta is essential for TCR-mediated T-cell activation, but is dispensable during TCR-dependent thymocyte development. TCR-initiated NF-kappaB activation was absent from PKC-theta(-/-) mature T lymphocytes, but was intact in thymocytes. Activation of NF-kappaB by tumour-necrosis factor alpha and interleukin-1 was unaffected in the mutant mice. Although studies in T-cell lines had suggested that PKC-theta regulates activation of the JNK signalling pathway, induction of JNK was normal in T cells from mutant mice. These results indicate that PKC-theta functions in a unique pathway that links the TCR signalling complex to the activation of NF-kappaB in mature T lymphocytes.

    View details for Web of Science ID 000086119000054

    View details for PubMedID 10746729