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401 Characterization of T-Cell Exhaustion in Rapid Progressing Multiple Myeloma Using Cross Center Scrna-Seq Study

Program: Oral and Poster Abstracts
Type: Oral
Session: 652. Multiple Myeloma and Plasma cell Dyscrasias: Clinical and Epidemiological: A Map for the Changing Landscape of Multiple Myeloma
Hematology Disease Topics & Pathways:
Translational Research
Sunday, December 12, 2021: 10:30 AM

William Pilcher, BSc1*, Beena E Thomas, Ph.D2,3*, Swati S Bhasin, PhD2,4, Reyka G Jayasinghe, PhD5,6*, Adeeb H Rahman, PhD7,8*, Seunghee Kim-Schulze, PhD9*, Edgar Gonzalez-Kozlova, PhD10*, Taxiarchis Kourelis, M.D.11, Madhav V Dhodapkar, MD12, Ravi Vij, MBBS13,14, Shaji K Kumar, MD11,15, Shaadi Mehr, PhD16*, Mark Hamilton, PhD17*, Hearn Jay Cho, MD, PhD18,19, Daniel Auclair, PhD20, David Avigan, MD21, Sacha Gnjatic, PhD7,22*, Li Ding, PhD23* and Manoj Bhasin, PhD4

1Emory University, Wallace H. Coulter Department of Biomedical Engineering, Atlanta, GA
2Aflac Cancer and Blood Disorders Center, Children’s Healthcare of Atlanta, Atlanta, GA
3Emory School of Medicine, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA
4Department of Pediatrics, Emory University School of Medicine, Atlanta, GA
5McDonnell Genome Institute, Washington University School of Medicine, Saint Louis, MO
6Department of Medicine, Washington University School of Medicine, Saint Louis, MO
7Mount Sinai School of Medicine, New York, NY
8Human Immune Monitoring Center, Icahn School of Medicine at Mount Sinai, New York, NY
9Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY
10Department of Genetics and Genomics., Icahn School of Medicine at Mount Sinai, New York
11Division of Hematology, Mayo Clinic, Rochester, MN
12Winship Cancer Institute, Emory University, Atlanta, GA
13Washington University School of Medicine, Ballwin, MO
14Division of Medical Oncology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
15Mayo Clinic, Rochester, MN
16Multiple Myeloma Research Foundation, Norwalk, CT
17The Multiple Myeloma Research Foundation (MMRF), Norwalk, CT
18Department of Medicine, Hematology and Medical Oncology, Icahn School of Medicine at Mount Sinai, New York, NY
19MMRF; Icahn School of Medicine at Mount Sinai, Norwalk, CT
20Multiple Myeloma Research Foundation (MMRF), Norwalk, CT
21Beth Israel Deaconess Medical Center, Boston, MA
22Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY
23Siteman Cancer Center, Washington University School of Medicine, Saint Louis, MO

Introduction: Multiple myeloma (MM) is a complex hematological malignancy with the heterogenous immune bone marrow (BM) environment contributing to tumor growth, drug resistance, and immune escape. T-Cells play a critical role in the clearance of malignant plasma cells from the tumor environment. However, T-Cells in multiple myeloma demonstrate impaired cytotoxicity, proliferation, and cytokine production due to the activation of immune inhibitory receptors from ligands produced by the myeloma cells. In this study, we investigate the behavior of T-Cells in MM patients by using single-cell RNA-Seq (scRNA-Seq) to compare the transcriptomic profiles of BM T-Cells of patients with rapid progressing (FP; PFS < 18mo) and non-progressing (NP; PFS > 4yrs) disease.

Methods: Newly diagnosed MM patients (n=18) from the Multiple Myeloma Research Foundation (MMRF) CoMMpass study (NCT01454297) were identified as either rapid progressors or non-progressors based on their progression free survival since diagnosis. To capture transcriptomic data, scRNA-Seq was performed on 48 aliquots of frozen CD138-negative BM cells at three medical centers/universities (Beth Israel Deaconess Medical Center, Boston, Washington University in St. Louis, and Mount Sinai School of Medicine, NYC). Samples were collected at diagnosis prior to treatment. Surface marker expression for 29 proteins was captured for at least one sample per patient using CITE-Seq. After integration and batch correction, clustering was performed to identify cells of T or NK lineage. Uniform Manifold Approximation and Projection (UMAP) and differential expression were used to identify T-Lymphoid subtypes, and differences in NP and FP samples.

Results: In this study, single cell transcriptomic profiles were identified for ~102,207 cells from 48 samples of 18 MM patients. 40,328 T (CD3+) and NK (CD3-, NKG7+) cells were isolated, and subclustered for further analysis (Fig 1A). Using differentially expressed markers for each cluster, the T-Lymphoid subset was refined into seven subtypes, consisting of various CD4+ T-Cells, CD8+ T-Cells, and NK cells (Fig 1B). The CD8+ cells were divided into three distinct phenotypes, namely a GZMK-, GZMB- CD8+ T-Cell cluster, a GZMK+ CD8+ Exhausted T-Cell cluster enriched in TIGIT and multiple chemokines (CCL3, CCL4, XCL2), and a GZMB+ NkT cluster enriched in cytolytic markers (PRF1, GNLY, NKG7) (Fig 1C). Differential expression between NP and FP samples in this CD8+ subset showed enrichment of the NkT cytotoxic markers in NP samples, while FP samples were enriched in the CD8+ Exhausted chemokine markers (Fig 1D). Furthermore, the proportion of CD8+ Exhausted T-Cells was enriched in FP samples (p.val < 0.05) (Fig 1E). Exhaustion markers were measured through both RNA and surface marker levels. In RNA, TIGIT was uniquely associated with the FP-enriched CD8+ Exhausted T-Cell cluster, and CD160 was uniquely expressed in FP samples (Fig 1F). CITE-Seq surface marker expression confirms enrichment of both TIGIT and PD1 in the CD8+ Exhausted T-Cell cluster, and along with more exhaustion in FP samples (p.val < 0.01).

Conclusion: In this study, we have identified significant differences in T-Cell activity in patients with non-progressing and rapid-progressing multiple myeloma. T-Cells in rapid progressing patients appear to be in a suppressed state, with low cytolytic activity and enriched exhaustion markers. This GZMK+ T-Cell population shows strong similarities with an aging-associated subtype of effector memory T-Cells found to be enriched in older populations (Mogilenko et al, Immunity 54, 2021). These findings will be further validated in an expanded study, consisting both of a larger number of samples, and multiple samples at different timepoints from the same patient.

Disclosures: Jayasinghe: MMRF: Consultancy; WUGEN: Consultancy. Vij: BMS: Research Funding; Takeda: Honoraria, Research Funding; Sanofi: Honoraria, Research Funding; BMS: Honoraria; GSK: Honoraria; Oncopeptides: Honoraria; Karyopharm: Honoraria; CareDx: Honoraria; Legend: Honoraria; Biegene: Honoraria; Adaptive: Honoraria; Harpoon: Honoraria. Kumar: Antengene: Consultancy, Honoraria; KITE: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Oncopeptides: Consultancy; Tenebio: Research Funding; Beigene: Consultancy; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Carsgen: Research Funding; Amgen: Consultancy, Research Funding; Merck: Research Funding; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Abbvie: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Consultancy, Research Funding; Novartis: Research Funding; Astra-Zeneca: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche-Genentech: Consultancy, Research Funding; Bluebird Bio: Consultancy; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Adaptive: Membership on an entity's Board of Directors or advisory committees, Research Funding; Sanofi: Research Funding. Avigan: Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Pharmacyclics: Research Funding; Kite Pharma: Consultancy, Research Funding; Juno: Membership on an entity's Board of Directors or advisory committees; Partner Tx: Membership on an entity's Board of Directors or advisory committees; Karyopharm: Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Aviv MedTech Ltd: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees; Legend Biotech: Membership on an entity's Board of Directors or advisory committees; Chugai: Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy; Parexcel: Consultancy; Takeda: Consultancy; Sanofi: Consultancy. Bhasin: Sanofi Genzyme: Consultancy; Canomiks: Membership on an entity's Board of Directors or advisory committees; Anxomics: Membership on an entity's Board of Directors or advisory committees.

*signifies non-member of ASH