Therapeutic Targeting of Mesothelin with Chimeric Antigen Receptor T Cells in Acute Myeloid Leukemia

Purpose: We previously identified mesothelin (MSLN) as highly expressed in a significant fraction of acute myeloid leukemia (AML) but entirely silent in normal hematopoiesis, providing a promising antigen for immunotherapeutic targeting that avoids hematopoietic toxicity. Given that T cells genetically modified to express chimeric antigen receptors (CAR) are effective at eradicating relapsed/refractory acute lymphocytic leukemia, we developed MSLN-directed CAR T cells for preclinical evaluation in AML. Experimental Design: The variable light (VL) and heavy (VH) sequences from the MSLN-targeting SS1P immunotoxin were used to construct the single-chain variable fragment of the standard CAR containing 41-BB costimulatory and CD3Zeta stimulatory domains. The preclinical efficacy of MSLN CAR T cells was evaluated against AML cell lines and patient samples expressing various levels of MSLN in vitro and in vivo. Results: We demonstrate that MSLN is expressed on the cell surface of AML blasts and leukemic stem cell–enriched CD34+CD38− subset, but not on normal hematopoietic stem and progenitor cells (HSPC). We further establish that MSLN CAR T cells are highly effective in eliminating MSLN-positive AML cells in cell line– and patient-derived xenograft models. Importantly, MSLN CAR T cells can target and eradicate CD34+CD38− cells without impacting the viability of normal HSPCs. Finally, we show that CAR T-cell functionality can be improved by inhibition of the ADAM17 metalloprotease that promotes shedding of MSLN. Conclusions: These findings demonstrate that MSLN is a viable target for CAR T-cell therapy in AML and that inhibiting MSLN shedding is a promising approach to improve CAR T-cell efficacy.

[1]  R. Ries,et al.  Mesothelin is a novel cell surface disease marker and potential therapeutic target in acute myeloid leukemia. , 2021, Blood advances.

[2]  R. Ries,et al.  Mesothelin Expression Is Associated with Extramedullary Disease and Promotes In Vivo Leukemic Growth in Acute Myeloid Leukemia , 2020 .

[3]  J. Pérez-Simón,et al.  Overcoming Chimeric Antigen Receptor (CAR) Modified T-Cell Therapy Limitations in Multiple Myeloma , 2020, Frontiers in Immunology.

[4]  S. Gill,et al.  CAR T Cells for Acute Myeloid Leukemia: State of the Art and Future Directions , 2020, Frontiers in Oncology.

[5]  He Huang,et al.  Successful chimeric antigen receptor T cells therapy in extramedullary relapses of acute lymphoblastic leukemia after allogeneic hematopoietic stem cell transplantation , 2019, Bone Marrow Transplantation.

[6]  R. Brentjens,et al.  Engineering strategies to overcome the current roadblocks in CAR T cell therapy , 2019, Nature Reviews Clinical Oncology.

[7]  D. Torigian,et al.  Phase I Study of Lentiviral-Transduced Chimeric Antigen Receptor-Modified T Cells Recognizing Mesothelin in Advanced Solid Cancers. , 2019, Molecular therapy : the journal of the American Society of Gene Therapy.

[8]  Yue Qin,et al.  Diagnostic value of soluble mesothelin-related peptides in pleural effusion for malignant pleural mesothelioma , 2019, Medicine.

[9]  I. Weissman,et al.  Surgical adhesions in mice are derived from mesothelial cells and can be targeted by antibodies against mesothelial markers , 2018, Science Translational Medicine.

[10]  R. Fluhrer,et al.  Proteolytic ectodomain shedding of membrane proteins in mammals—hardware, concepts, and recent developments , 2018, The EMBO journal.

[11]  Michel Sadelain,et al.  Chimeric Antigen Receptor Therapy. , 2018, The New England journal of medicine.

[12]  D. Torigian,et al.  Activity of Mesothelin-Specific Chimeric Antigen Receptor T Cells Against Pancreatic Carcinoma Metastases in a Phase 1 Trial. , 2018, Gastroenterology.

[13]  Michael L. Wang,et al.  T Cells Genetically Modified to Express an Anti-B-Cell Maturation Antigen Chimeric Antigen Receptor Cause Remissions of Poor-Prognosis Relapsed Multiple Myeloma. , 2018, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[14]  R. Hagedoorn,et al.  A Jurkat 76 based triple parameter reporter system to evaluate TCR functions and adoptive T cell strategies , 2018, Oncotarget.

[15]  N. Heussen,et al.  Influence of MMP inhibitor GM6001 loading of fibre coated polypropylene meshes on wound healing: Implications for hernia repair , 2018, Journal of biomaterials applications.

[16]  Yang Feng,et al.  CD22-CAR T Cells Induce Remissions in CD19-CAR Naïve and Resistant B-ALL , 2017, Nature Medicine.

[17]  S. Jagannath,et al.  Durable Clinical Responses in Heavily Pretreated Patients with Relapsed/Refractory Multiple Myeloma: Updated Results from a Multicenter Study of bb2121 Anti-Bcma CAR T Cell Therapy , 2017 .

[18]  P. Baldo,et al.  Amatuximab and novel agents targeting mesothelin for solid tumors , 2017, OncoTargets and therapy.

[19]  I. Pastan,et al.  Panbinostat decreases cFLIP and enhances killing of cancer cells by immunotoxin LMB-100 by stimulating the extrinsic apoptotic pathway , 2017, Oncotarget.

[20]  Ø. Bruserud,et al.  Therapeutic targeting of leukemic stem cells in acute myeloid leukemia – the biological background for possible strategies , 2017, Expert opinion on drug discovery.

[21]  I. Pastan,et al.  Mesothelin Immunotherapy for Cancer: Ready for Prime Time? , 2016, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  S. Heimfeld,et al.  Immunotherapy of non-Hodgkin’s lymphoma with a defined ratio of CD8+ and CD4+ CD19-specific chimeric antigen receptor–modified T cells , 2016, Science Translational Medicine.

[23]  Daniel Li,et al.  CD19 CAR-T cells of defined CD4+:CD8+ composition in adult B cell ALL patients. , 2016, The Journal of clinical investigation.

[24]  M. I. Rosa,et al.  Mesothelin as a biomarker for ovarian carcinoma: a meta-analysis. , 2016, Anais da Academia Brasileira de Ciencias.

[25]  I. Pastan,et al.  Reduced Shedding of Surface Mesothelin Improves Efficacy of Mesothelin-Targeting Recombinant Immunotoxins , 2016, Molecular Cancer Therapeutics.

[26]  Yulei N. Wang,et al.  Phase I Study of DMOT4039A, an Antibody–Drug Conjugate Targeting Mesothelin, in Patients with Unresectable Pancreatic or Platinum-Resistant Ovarian Cancer , 2016, Molecular Cancer Therapeutics.

[27]  E. Jaffee,et al.  Safety and survival with GVAX pancreas prime and Listeria Monocytogenes-expressing mesothelin (CRS-207) boost vaccines for metastatic pancreatic cancer. , 2015, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[28]  I. Pastan,et al.  Methylation-Associated Partial Down-Regulation of Mesothelin Causes Resistance to Anti-Mesothelin Immunotoxins in a Pancreatic Cancer Cell Line , 2015, PloS one.

[29]  S. Meshinchi,et al.  Pediatric acute myeloid leukemia: biology and therapeutic implications of genomic variants. , 2015, Pediatric clinics of North America.

[30]  S. Riddell,et al.  The Nonsignaling Extracellular Spacer Domain of Chimeric Antigen Receptors Is Decisive for In Vivo Antitumor Activity , 2014, Cancer Immunology Research.

[31]  D. Torigian,et al.  Mesothelin-Specific Chimeric Antigen Receptor mRNA-Engineered T Cells Induce Antitumor Activity in Solid Malignancies , 2013, Cancer Immunology Research.

[32]  B. Levine,et al.  T Cells Expressing Chimeric Antigen Receptors Can Cause Anaphylaxis in Humans , 2013, Cancer Immunology Research.

[33]  B. Levine,et al.  Cells Expressing Chimeric Antigen Receptors Can Cause Anaphylaxis in Humans , 2013 .

[34]  I. Pastan,et al.  Antigen shedding may improve efficiencies for delivery of antibody-based anticancer agents in solid tumors. , 2012, Cancer research.

[35]  I. Pastan,et al.  Modulating Mesothelin Shedding to Improve Therapy , 2012, Oncotarget.

[36]  E. Jaffee,et al.  A Live-Attenuated Listeria Vaccine (ANZ-100) and a Live-Attenuated Listeria Vaccine Expressing Mesothelin (CRS-207) for Advanced Cancers: Phase I Studies of Safety and Immune Induction , 2011, Clinical Cancer Research.

[37]  I. Pastan,et al.  Cytotoxic activity of immunotoxin SS1P is modulated by TACE-dependent mesothelin shedding. , 2011, Cancer research.

[38]  B. Christensen,et al.  The relationship between tumor MSLN methylation and serum mesothelin (SMRP) in mesothelioma , 2011, Epigenetics.

[39]  C. Pui,et al.  Biology, risk stratification, and therapy of pediatric acute leukemias: an update. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[40]  P. Johnston,et al.  Oncogenic Kras promotes chemotherapy-induced growth factor shedding via ADAM17. , 2011, Cancer research.

[41]  R. Arceci,et al.  Biology of Acute Myeloid Leukemia , 2011 .

[42]  P. Johnston,et al.  Chemotherapy-Induced Activation of ADAM-17: A Novel Mechanism of Drug Resistance in Colorectal Cancer , 2010, Clinical Cancer Research.

[43]  D. Doval,et al.  Clinical benefit of INCB7839, a potent and selective ADAM inhibitor, in combination with trastuzumab in patients with metastatic HER2+ breast cancer. , 2010 .

[44]  D. Doval,et al.  Clinical Benefit of INCB7839, a Potent and Selective Inhibitor of ADAM10 and ADAM17, in Combination with Trastuzumab in Metastatic HER2 Positive Breast Cancer Patients. , 2009 .

[45]  J. Fridman,et al.  Synergistic inhibition with a dual epidermal growth factor receptor/HER-2/neu tyrosine kinase inhibitor and a disintegrin and metalloprotease inhibitor. , 2008, Cancer research.

[46]  Mitchell Ho,et al.  Mesothelin Is Shed from Tumor Cells , 2006, Cancer Epidemiology Biomarkers & Prevention.

[47]  P. Stern,et al.  The Role of Extracellular Spacer Regions in the Optimal Design of Chimeric Immune Receptors: Evaluation of Four Different scFvs and Antigens , 2005, Journal of immunotherapy.

[48]  I. Pastan,et al.  Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity. , 1998, Proceedings of the National Academy of Sciences of the United States of America.