-Author name in bold denotes the presenting author
-Asterisk * with author name denotes a Non-ASH member
Clinically Relevant Abstract denotes an abstract that is clinically relevant.

PhD Trainee denotes that this is a recommended PHD Trainee Session.

Ticketed Session denotes that this is a ticketed session.

4796 Dissection of Single-Cell Landscapes for the Development of Chimeric Antigen Receptor T Cells in Hodgkin Lymphoma

Program: Oral and Poster Abstracts
Session: 702. CAR-T Cell Therapies: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Research, Translational Research
Monday, December 9, 2024, 6:00 PM-8:00 PM

Adrian Gottschlich, MD1,2,3,4*, Ruth Grünmeier2*, Gordon Victor Hoffmann2*, Sayantan Nandi, PhD2*, Vladyslav Kavaka, MD5,6*, Philipp Jie Müller2*, Jakob Jobst2*, Arman Öner, MD2*, Rainer Kaiser, MD7,8*, Jan Gärtig2*, Ignazio Piseddu, MD9,10*, Stephanie Frenz-Wiessner, MD11*, Heiko Schulz, MD12*, Lea Di Fina8*, Maité Mulkers8*, Moritz Thomas, PhD13,14*, Veronika Igl2*, Thomas Alexander Janert2*, Daria Briukhovetska, PhD2*, Donjetë Simnica, PhD2*, Emanuele Carlini2*, Christina Tsiverioti2*, Marcel Trefny, PhD2*, Theo Lorenzini, MD2*, Florian Märkl, PhD2*, Ruben Brabenec2,14*, Thaddäus Strzalkowski2*, Sophia Stock, MD2,3,15*, Stefanos Michaelides2*, Irmela Jeremias, MD3,11, Johannes Christian Hellmuth, MD16*, Martin Thelen17,18*, Sarah Reinke, PhD19*, Wolfram Klapper, MD, Prof19*, Pascal Gelebart, PhD20,21*, Leo Nicolai, MD7,8*, Carsten Marr, PhD14*, Eduardo Beltrán, PhD5,6,22*, Christoph Klein, MD9,11, Fanny Baran-Marszak, MD, PhD23,24*, Paul J. Bröckelmann, MD25*, Andreas Rosenwald, MD26,27*, Michael von Bergwelt-Baildon, MD, PhD1,3,4*, Stefan Endres, MD, Prof2,3,28* and Sebastian Kobold, MD2,3,28*

1Department of Medicine III, University Hospital, LMU Munich, Munich, Germany
2Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany
3German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany
4Bavarian Cancer Research Center (BZKF), Munich, Germany
5Institute of Clinical Neuroimmunology, University Hospital, LMU Munich, Munich, Germany
6Biomedical Center (BMC), Faculty of Medicine, LMU Munich, Martinsried, Germany
7DZHK (German Center for Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany
8Department of Medicine I, University Hospital, LMU Munich, Munich, Germany
9Gene Center and Department of Biochemistry, LMU Munich, Munich, Germany
10LMU University Hospital, LMU Munich, Department of Medicine II, Munich, Germany
11Department of Pediatrics, Dr. von Hauner Children’s Hospital, University Hospital, LMU Munich, Munich, Germany
12Institute of Pathology, University Hospital, LMU Munich, Munich, DEU
13School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
14Institute of AI for Health, Helmholtz Zentrum München - German Research Center for Environmental Health, Neuherberg, Germany
15Department of Medicine III, LMU University Hospital, LMU Munich, Munich, Germany
16Department of Medicine III, LMU Munich, Munich, Germany
17Department of General, Visceral, Cancer and Transplantation Surgery, University of Cologne, Cologne, DEU
18Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany
19Department of Pathology, Hematopathology Section, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany
20Department of Clinical Science, University of Bergen, Bergen, NOR
21Department of Hematology, Haukeland University Hospital, Bergen, Norway
22Munich Cluster for Systems Neurology (SyNergy), Munich, Germany
23Service d’Hématologie Biologique, Hôpitaux Universitaire Paris Seine Saint Denis (HUPSSD), Hôpital Avicenne, Université Sorbonne Paris Nord, Bobigny, France
24INSERM U978, University of Paris 13, Bobigny, France
25University of Cologne, Faculty of Medicine and University Hospital of Cologne, Department I of Internal Medicine, and Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf (CIO ABCD), and German Hodgkin Study Group (GHSG), Cologne, Germany
26Comprehensive Cancer Center Mainfranken, University Hospital Würzburg, Würzburg, Germany
27Institute of Pathology, Julius-Maximilians-Universität Würzburg, Würzburg, Germany
28Einheit für Klinische Pharmakologie (EKLiP), Helmholtz Zentrum München, German Research Center for Environmental Health (HMGU), Neuherberg, Germany

The success of targeted immunotherapies for hematological malignancies has heralded their potential as salvage therapies as well as in earlier treatment lines (Cappell & Kochenderfer, 2023). While conventional chemotherapy-based treatments can achieve long-term survival in up to 90 % of treated patients with classic Hodgkin lymphoma (cHL), these therapies are associated with treatment-related comorbidities, calling for more tailored and specific approaches (Schaapveld et al., 2015; Shanbhag & Ambinder, 2018). While targeted treatments, especially immunotherapies are taking oncology by storm, the utility in cHL is so far limited to CD30 and PD-1-targeting strategies and there is a clear lack of drugable relevant target structures in this disease. This can be partly attributed to technical difficulties of analyzing the malignant Hodgkin-Reed-Sternberg (HRS) cells specifically. Capitalizing on our previous work using large scale data mining to inform target discovery, we hypothesized that combining different analytical methods with large single-cell RNA-Sequencing (scRNA-Seq) datasets would permit selective target definition with functional relevance to the disease and thereby allow the development of novel immunotherapeutic strategies.

Leveraging microarray profiles of laser-dissected HRS cells and a scRNA-Seq cohort of cHL patients (total of n = 44 primary samples; n = 34 cHL samples; n = 10 RLN (reactive lymph node) control samples), we screened for novel target antigens highly expressed on HRS cells with functional relevance in the tumor microenvironement (TME) of cHL. Unbiased in silico analyses revealed CD80, CD86 and PD-L1 as most suitable candidate target antigens with CD86 showing the highest expression on HRS cells. ScRNA-Seq analyses unveiled a shift of the CD80-CD86-CTLA-4-CD28 towards the immunosuppressive CTLA-4 axis in the TME of cHL compared to RLN controls. In advanced cell culture models, including iPSC-derived organoid models, blockage of CD86 lead to the decreased expression of PD-1 and CTLA-4 and an overall reversal of the exhaustive phenotype of cHL-associated T cells. High protein expression of CD86 on HRS cells and in the TME (cHL-infiltrating tumor-associated macrophages (cHL-TAM), B cells) was confirmed in different validation cohorts including relapsed and refractory cHL (r/r cHL) patients by conventional immunohistochemistry and multiplexed immunofluorescence (n = 34 cHL patients). Following target identification, CAR T cells redirected against CD86 were developed and the functionality of these CAR T cells was investigated in preclinical models both in vitro and in vivo. Anti‑CD86 CAR T cells effectively deplete cHL-TAM and are highly effective in various in vitro and in vivo models of cHL, including models of CD30-negative disease.

Given the fundamental role of the CD80-CD86-CTLA-4-CD28 axis in the generation of the adaptive immune response, detailed toxicity assessments were carried out leveraging murine surrogate anti-CD86 CAR T cells, with similar binding and activation thresholds as their human counterpart. These anti-mCD86 CAR T cells did not cause toxicities in lymphodepleted, immunocompetent mice. In addition, the impact of anti-CD86-directed immunotherapies (e.g. anti-CD86-blocking antibodies, anti-mCD86 CAR T cells) on bacterial host defense and formation of antigen-specific adaptive immunity was investigated in syngeic mouse models. Anti-CD86 immunotherapy did not lead to enhanced bacteremia in a model of gram-negative sepsis, while preclinical vaccination models revealed a mildy reduced formation of antigen-specific T cell development in mice.

In summary, we provide a framework for unbiased, multi-dimensional target screening and highlight the functional relevance of the immunosuppressive CD86-CTLA-4 axis in cHL. CD86-directed immunotherapy could reverse the exhaustive phenotype of cHL-associated T cells, while demonstrating strong treatment efficacy in xenograft mouse models. Importantly, elaborate toxicity assessments of anti-CD86-targeted immunotherapies utilizing syngenic mouse models did not reveal measureable toxicity in mice. Overall, our data emphasizes the vast translational potential of CD86-targeted immunotherapies in cHL and provide a strong rationale for further clinical investigations.

Disclosures: Gottschlich: Nanogami: Research Funding; Tabby Therapeutics: Research Funding. Jeremias: Tubulis GmbH: Patents & Royalties: pending patent application FLT3-mAb 20D9. Klapper: Roche, Janssen, Amgen, InCyte: Research Funding. Bröckelmann: Else-Kröner Fresenius Foundation: Other: Excellence Stipend; Takeda: Consultancy, Honoraria, Research Funding; Stemline: Consultancy, Honoraria; Need Inc.: Consultancy, Current holder of stock options in a privately-held company; Merck Sharp & Dohme: Consultancy, Honoraria, Research Funding; BMS: Honoraria, Research Funding; BeiGene: Honoraria, Research Funding. von Bergwelt-Baildon: AMGEN, Astellas, AstraZeneca, Bristol-Myers Squibb, Daiichi Sankyo, KITE/Gilead Mologen, Miltenyi, MSD Sharp + Dohme, Novartis, Priothera, Roche, TABBY: Consultancy, Honoraria, Research Funding, Speakers Bureau; TABBY: Membership on an entity's Board of Directors or advisory committees. Endres: TCR2 Inc: Other: Licence fees, Research Funding; Carina Biotech: Other: Licence Fees; Arcus Bioscience: Research Funding; Catalym GmbH: Research Funding; Plectonic GmbH: Research Funding. Kobold: CR2 Inc., Miltenyi, Galapagos, Novartis, BMS, and GS: Honoraria; CR2 Inc and Carina Biotech: Other: Licence fees; CR2 Inc., Tabby Therapeutics, Catalym GmBH, Plectonic GmBH and Arcus Bioscience: Research Funding.

*signifies non-member of ASH