Tassilo Leo Alexander Wachsmann

All Publications

• Targeting epigenetic regulation and post-translational modification with 5-Aza-2' deoxycytidine and SUMO E1 inhibition augments T-cell receptor therapy JOURNAL FOR IMMUNOTHERAPY OF CANCER Kroonen, J. S., Wouters, A. K., de Graaf, I. J., Remst, D. G., Kumar, S., Wachsmann, T. A., Teunisse, A. S., Roelands, J. P., de Miranda, N. C., Griffioen, M., Heemskerk, M. M., Vertegaal, A. O. 2024; 12 (9)

  Abstract

  Cellular immunotherapy using modified T cells offers new avenues for cancer treatment. T-cell receptor (TCR) engineering of CD8 T cells enables these cells to recognize tumor-associated antigens and tumor-specific neoantigens. Improving TCR T-cell therapy through increased potency and in vivo persistence will be critical for clinical success.We evaluated a novel drug combination to enhance TCR therapy in mouse models for acute myeloid leukemia (AML) and multiple myeloma (MM).Combining TCR therapy with the SUMO E1 inhibitor TAK981 and the DNA methylation inhibitor 5-Aza-2' deoxycytidine resulted in strong antitumor activity in a persistent manner against two in vivo tumor models of established AML and MM. We uncovered that the drug combination caused strong T-cell proliferation, increased cytokine signaling in T cells, improved persistence of T cells, and reduced differentiation towards exhausted phenotype. Simultaneously the drug combination enhanced immunogenicity of the tumor by increasing HLA and co-stimulation and surprisingly reducing inhibitory ligand expression.Combining T-cell therapy with TAK981 and 5-Aza-2' deoxycytidine may be an important step towards improved clinical outcome.

  View details for DOI 10.1136/jitc-2023-008654

  View details for Web of Science ID 001322016100001

  View details for PubMedID 39326886

  View details for PubMedCentralID PMC11425949

• Combining BCMA-targeting CAR T cells with TCR-engineered T-cell therapy to prevent immune escape of multiple myeloma BLOOD ADVANCES Wachsmann, T. A., Meeuwsen, M. H., Remst, D. G., Buchner, K., Wouters, A. K., Hagedoorn, R. S., Falkenburg, J., Heemskerk, M. M. 2023; 7 (20): 6178-6183

  View details for DOI 10.1182/bloodadvances.2023010410

  View details for Web of Science ID 001097889000001

  View details for PubMedID 37567150

  View details for PubMedCentralID PMC10582830

• High-affinity CD8 variants enhance the sensitivity of pMHCI<i> via</i> low TCRs JOURNAL OF BIOLOGICAL CHEMISTRY Knezevic, L., Wachsmann, T. A., Francis, O., Dockree, T., Bridgeman, J. S., Wouters, A., de Wet, B., Cole, D. K., Clement, M., McLaren, J. E., Gostick, E., Ladell, K., Llewellyn-Lacey, S., Price, D. A., van den Berg, H. A., Tabi, Z., Sessions, R. B., Heemskerk, M. M., Wooldridge, L. 2023; 299 (8): 104981

  Abstract

  CD8+ T cell-mediated recognition of peptide-major histocompatibility complex class I (pMHCI) molecules involves cooperative binding of the T cell receptor (TCR), which confers antigen specificity, and the CD8 coreceptor, which stabilizes the TCR/pMHCI complex. Earlier work has shown that the sensitivity of antigen recognition can be regulated in vitro by altering the strength of the pMHCI/CD8 interaction. Here, we characterized two CD8 variants with moderately enhanced affinities for pMHCI, aiming to boost antigen sensitivity without inducing non-specific activation. Expression of these CD8 variants in model systems preferentially enhanced pMHCI antigen recognition in the context of low-affinity TCRs. A similar effect was observed using primary CD4+ T cells transduced with cancer-targeting TCRs. The introduction of high-affinity CD8 variants also enhanced the functional sensitivity of primary CD8+ T cells expressing cancer-targeting TCRs, but comparable results were obtained using exogenous wild-type CD8. Specificity was retained in every case, with no evidence of reactivity in the absence of cognate antigen. Collectively, these findings highlight a generically applicable mechanism to enhance the sensitivity of low-affinity pMHCI antigen recognition, which could augment the therapeutic efficacy of clinically relevant TCRs.

  View details for DOI 10.1016/j.jbc.2023.104981

  View details for Web of Science ID 001166350400001

  View details for PubMedID 37390984

  View details for PubMedCentralID PMC10432799

• PRAME and CTCFL-reactive TCRs for the treatment of ovarian cancer FRONTIERS IN IMMUNOLOGY van Amerongen, R. A., Tuit, S., Wouters, A. K., van de Meent, M., Siekman, S. L., Meeuwsen, M. H., Wachsmann, T. A., Remst, D. G., Hagedoorn, R. S., van der Steen, D. M., de Ru, A. H., Verdegaal, E. E., van Veelen, P. A., Falkenburg, J., Heemskerk, M. M. 2023; 14: 1121973

  Abstract

  Recurrent disease emerges in the majority of patients with ovarian cancer (OVCA). Adoptive T-cell therapies with T-cell receptors (TCRs) targeting tumor-associated antigens (TAAs) are considered promising solutions for less-immunogenic 'cold' ovarian tumors. In order to treat a broader patient population, more TCRs targeting peptides derived from different TAAs binding in various HLA class I molecules are essential. By performing a differential gene expression analysis using mRNA-seq datasets, PRAME, CTCFL and CLDN6 were selected as strictly tumor-specific TAAs, with high expression in ovarian cancer and at least 20-fold lower expression in all healthy tissues of risk. In primary OVCA patient samples and cell lines we confirmed expression and identified naturally expressed TAA-derived peptides in the HLA class I ligandome. Subsequently, high-avidity T-cell clones recognizing these peptides were isolated from the allo-HLA T-cell repertoire of healthy individuals. Three PRAME TCRs and one CTCFL TCR of the most promising T-cell clones were sequenced, and transferred to CD8+ T cells. The PRAME TCR-T cells demonstrated potent and specific antitumor reactivity in vitro and in vivo. The CTCFL TCR-T cells efficiently recognized primary patient-derived OVCA cells, and OVCA cell lines treated with demethylating agent 5-aza-2'-deoxycytidine (DAC). The identified PRAME and CTCFL TCRs are promising candidates for the treatment of patients with ovarian cancer, and are an essential addition to the currently used HLA-A*02:01 restricted PRAME TCRs. Our selection of differentially expressed genes, naturally expressed TAA peptides and potent TCRs can improve and broaden the use of T-cell therapies for patients with ovarian cancer or other PRAME or CTCFL expressing cancers.

  View details for DOI 10.3389/fimmu.2023.1121973

  View details for Web of Science ID 000962177900001

  View details for PubMedID 37026005

  View details for PubMedCentralID PMC10070997

• Broadly applicable TCR-based therapy for multiple myeloma targeting the immunoglobulin J chain JOURNAL OF HEMATOLOGY & ONCOLOGY Meeuwsen, M. H., Wouters, A. K., Wachsmann, T. A., Hagedoorn, R. S., Kester, M. D., Remst, D. G., van der Steen, D. M., de Ru, A. H., van Hees, E. P., Kremer, M., Griffioen, M., van Veelen, P. A., Falkenburg, J., Heemskerk, M. M. 2023; 16 (1): 16

  Abstract

  The immunoglobulin J chain (Jchain) is highly expressed in the majority of multiple myeloma (MM), and Jchain-derived peptides presented in HLA molecules may be suitable antigens for T-cell therapy of MM.Using immunopeptidomics, we identified Jchain-derived epitopes presented by MM cells, and pHLA tetramer technology was used to isolate Jchain-specific T-cell clones.We identified T cells specific for Jchain peptides presented in HLA-A1, -A24, -A3, and -A11 that recognized and lysed JCHAIN-positive MM cells. TCRs of the most promising T-cell clones were sequenced, cloned into retroviral vectors, and transferred to CD8 T cells. Jchain TCR T cells recognized target cells when JCHAIN and the appropriate HLA restriction alleles were expressed, while JCHAIN or HLA-negative cells, including healthy subsets, were not recognized. Patient-derived JCHAIN-positive MM samples were also lysed by Jchain TCR T cells. In a preclinical in vivo model for established MM, Jchain-A1, -A24, -A3, and -A11 TCR T cells strongly eradicated MM cells, which resulted in 100-fold lower tumor burden in Jchain TCR versus control-treated mice.We identified TCRs targeting Jchain-derived peptides presented in four common HLA alleles. All four TCRs demonstrated potent preclinical anti-myeloma activity, encouraging further preclinical testing and ultimately clinical development.

  View details for DOI 10.1186/s13045-023-01408-6

  View details for Web of Science ID 000941154700001

  View details for PubMedID 36850001

  View details for PubMedCentralID PMC9969645

• Comparing CAR and TCR engineered T cell performance as a function of tumor cell exposure ONCOIMMUNOLOGY Wachsmann, T. A., Wouters, A. K., Remst, D. G., Hagedoorn, R. S., Meeuwsen, M. H., van Diest, E., Leusen, J., Kuball, J., Falkenburg, J., Heemskerk, M. M. 2022; 11 (1): 2033528

  Abstract

  Chimeric antigen receptor (CAR) T cell therapies have resulted in profound clinical responses in the treatment of CD19-positive hematological malignancies, but a significant proportion of patients do not respond or relapse eventually. As an alternative to CAR T cells, T cells can be engineered to express a tumor-targeting T cell receptor (TCR). Due to HLA restriction of TCRs, CARs have emerged as a preferred treatment moiety when targeting surface antigens, despite the fact that functional differences between engineered TCR (eTCR) T and CAR T cells remain ill-defined. Here, we compared the activity of CAR T cells versus engineered TCR T cells in targeting the B cell malignancy-associated antigen CD20 as a function of antigen exposure. We found CAR T cells to be more potent effector cells, producing higher levels of cytokines and killing more efficiently than eTCR T cells in a short time frame. However, we revealed that the increase of antigen exposure significantly impaired CAR T cell expansion, a phenotype defined by high expression of coinhibitory molecules and effector differentiation. In contrast, eTCR T cells expanded better than CAR T cells under high antigenic pressure, with lower expression of coinhibitory molecules and maintenance of an early differentiation phenotype, and comparable clearance of tumor cells.

  View details for DOI 10.1080/2162402X.2022.2033528

  View details for Web of Science ID 000749319100001

  View details for PubMedID 35127255

  View details for PubMedCentralID PMC8812760

• A library of cancer testis specific T cell receptors for T cell receptor gene therapy MOLECULAR THERAPY ONCOLYTICS de Rooij, M. J., Remst, D. G., van der Steen, D. M., Wouters, A. K., Hagedoorn, R. S., Kester, M. D., Meeuwsen, M. H., Wachsmann, T. A., de Ru, A. H., van Veelen, P. A., Verdegaal, E. E., Falkenburg, J., Heemskerk, M. M. 2023; 28: 1-14

  Abstract

  To increase the number of cancer patients that can be treated with T cell receptor (TCR) gene therapy, we aimed to identify a set of high-affinity cancer-specific TCRs targeting different melanoma-associated antigens (MAGEs). In this study, peptides derived from MAGE genes with tumor-specific expression pattern were identified by human leukocyte antigen (HLA) peptidomics. Next, peptide-HLA tetramers were generated, and used to sort MAGE-specific CD8+ T cell clones from the allogeneic (allo) HLA repertoire of healthy donors. To evaluate the clinical potential, most potent TCRs were sequenced, transferred into peripheral blood-derived CD8+ T cells, and tested for antitumor efficacy. In total we identified, seven MAGE-specific TCRs that effectively target MAGE-A1, MAGE-A3, MAGE-A6, and MAGE-A9 in the context of HLA-A∗01:01, -A∗02:01, -A∗03:01, -B∗07:02, -B∗35:01, or -C∗07:02. TCR gene transfer into CD8⁺ T cells resulted in efficient reactivity against a variety of different tumor types, while no cross-reactivity was detected. In addition, major in vivo antitumor effects of MAGE-A1 specific TCR engineered CD8⁺ T cells were observed in the orthotopic xenograft model for established multiple myeloma. The identification of seven MAGE-specific TCRs expands the pool of cancer patients eligible for TCR gene therapy and increases possibilities for personalized TCR gene therapy.

  View details for DOI 10.1016/j.omto.2022.11.007

  View details for Web of Science ID 000906126700001

  View details for PubMedID 36589698

  View details for PubMedCentralID PMC9792401

• T cell receptor engineering of primary NK cells to therapeutically target tumors and tumor immune evasion JOURNAL FOR IMMUNOTHERAPY OF CANCER Morton, L. T., Wachsmann, T. A., Meeuwsen, M. H., Wouters, A. K., Remst, D. G., van Loenen, M. M., Falkenburg, J., Heemskerk, M. M. 2022; 10 (3)

  Abstract

  T cell receptor (TCR)-engineered cells can be powerful tools in the treatment of malignancies. However, tumor resistance by Human Leukocyte antigen (HLA) class I downregulation can negatively impact the success of any TCR-mediated cell therapy. Allogeneic natural killer (NK) cells have demonstrated efficacy and safety against malignancies without inducing graft-versus-host-disease, highlighting the feasibility for an 'off the shelf' cellular therapeutic. Furthermore, primary NK cells can target tumors using a broad array of intrinsic activation mechanisms. In this study, we combined the antitumor effector functions of NK cells with TCR engineering (NK-TCR), creating a novel therapeutic strategy to avoid TCR-associated immune resistance.BOB1, is a transcription factor highly expressed in all healthy and malignant B cell lineages, including multiple myeloma (MM). Expression of an HLA-B*07:02 restricted BOB1-specifc TCR in peripheral blood-derived NK cells was achieved following a two-step retroviral transduction protocol. NK-TCR was then compared with TCR-negative NK cells and CD8-T cells expressing the same TCR for effector function against HLA-B*07:02+ B-cell derived lymphoblastoid cell lines (B-LCL), B-cell acute lymphoblastic leukemia and MM cell lines in vitro and in vivo.Firstly, TCR could be reproducibly expressed in NK cells isolated from the peripheral blood of multiple healthy donors generating pure NK-TCR cell products. Secondly, NK-TCR demonstrated antigen-specific effector functions against malignancies which were previously resistant to NK-mediated lysis and enhanced NK efficacy in vivo using a preclinical xenograft model of MM. Moreover, antigen-specific cytotoxicity and cytokine production of NK-TCR was comparable to CD8 T cells expressing the same TCR. Finally, in a model of HLA-class I loss, tumor cells with B2M KO were lysed by NK-TCR in an NK-mediated manner but were resistant to T-cell based killing.NK-TCR cell therapy enhances NK cell efficacy against tumors through additional TCR-mediated lysis. Furthermore, the dual efficacy of NK-TCR permits the specific targeting of tumors and the associated TCR-associated immune resistance, making NK-TCR a unique cellular therapeutic.

  View details for DOI 10.1136/jitc-2021-003715

  View details for Web of Science ID 000769335300007

  View details for PubMedID 35288464

  View details for PubMedCentralID PMC8921915

• An HLA-A*11:01-Binding Neoantigen from Mutated NPM1 as Target for TCR Gene Therapy in AML CANCERS van der Lee, D. I., Koutsoumpli, G., Reijmers, R. M., Honders, M., de Jong, R. M., Remst, D. G., Wachsmann, T. A., Hagedoorn, R. S., Franken, K. C., Kester, M. D., Harber, K. J., Roelofsen, L. M., Schouten, A. M., Mulder, A., Drijfhout, J. W., Veelken, H., van Veelen, P. A., Heemskerk, M. M., Falkenburg, J., Griffioen, M. 2021; 13 (21)

  Abstract

  Acute myeloid leukemia (AML) is a hematological malignancy caused by clonal expansion of myeloid progenitor cells. Most patients with AML respond to chemotherapy, but relapses often occur and infer a very poor prognosis. Thirty to thirty-five percent of AMLs carry a four base pair insertion in the nucleophosmin 1 gene (NPM1) with a C-terminal alternative reading frame of 11 amino acids. We previously identified various neopeptides from the alternative reading frame of mutant NPM1 (dNPM1) on primary AML and isolated an HLA-A*02:01-restricted T-cell receptor (TCR) that enables human T-cells to kill AML cells upon retroviral gene transfer. Here, we isolated T-cells recognizing the dNPM1 peptide AVEEVSLRK presented in HLA-A*11:01. The TCR cloned from a T-cell clone recognizing HLA-A*11:01+ primary AML cells conferred in vitro recognition and lysis of AML upon transfer to CD8 cells, but failed to induce an anti-tumor effect in immunodeficient NSG mice engrafted with dNPM1 OCI-AML3 cells. In conclusion, our data show that AVEEVSLRK is a dNPM1 neoantigen on HLA-A*11:01+ primary AMLs. CD8 cells transduced with an HLA-A*11:01-restricted TCR for dNPM1 were reactive against AML in vitro. The absence of reactivity in a preclinical mouse model requires further preclinical testing to predict the potential efficacy of this TCR in clinical development.

  View details for DOI 10.3390/cancers13215390

  View details for Web of Science ID 000726594700001

  View details for PubMedID 34771556

  View details for PubMedCentralID PMC8582585

• Protein kinase A regulates inflammatory pain sensitization by modulating HCN2 channel activity in nociceptive sensory neurons PAIN Herrmann, S., Rajab, H., Christ, I., Schirdewahn, C., Hoefler, D., Fischer, M. M., Bruno, A., Fenske, S., Gruner, C., Kramer, F., Wachsmann, T., Wahl-Schott, C., Stieber, J., Biel, M., Ludwig, A. 2017; 158 (10): 2012-2024

  Abstract

  Several studies implicated cyclic adenosine monophosphate (cAMP) as an important second messenger for regulating nociceptor sensitization, but downstream targets of this signaling pathway which contribute to neuronal plasticity are not well understood. We used a Cre/loxP-based strategy to disable the function of either HCN2 or PKA selectively in a subset of peripheral nociceptive neurons and analyzed the nociceptive responses in both transgenic lines. A near-complete lack of sensitization was observed in both mutant strains when peripheral inflammation was induced by an intradermal injection of 8br-cAMP. The lack of HCN2 as well as the inhibition of PKA eliminated the cAMP-mediated increase of calcium transients in dorsal root ganglion neurons. Facilitation of Ih via cAMP, a hallmark of the Ih current, was abolished in neurons without PKA activity. Collectively, these results show a significant contribution of both genes to inflammatory pain and suggest that PKA-dependent activation of HCN2 underlies cAMP-triggered neuronal sensitization.

  View details for DOI 10.1097/j.pain.0000000000001005

  View details for Web of Science ID 000412456000020

  View details for PubMedID 28767511