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172 Multi-Dimensional Molecular and Tumor-Microenvironment Analysis of Classic Hodgkin Lymphoma

Program: Oral and Poster Abstracts
Type: Oral
Session: 621. Lymphomas: Translational – Molecular and Genetic: Microenvironment and Methylome in Lymphoma and Misc.
Hematology Disease Topics & Pathways:
Research, Hodgkin lymphoma, adult, Translational Research, Lymphomas, genomics, Diseases, immunology, Lymphoid Malignancies, young adult , Biological Processes, Technology and Procedures, profiling, Study Population, Human, imaging
Saturday, December 9, 2023: 2:45 PM

Tomohiro Aoki, MD, PhD1,2, Gerben Duns, PhD3*, Shinya Rai, MD, PhD3*, Andrew Lytle, MD, PhD3, Yifan Yin3*, Clementine Sarkozy4*, Stacy Hung, PhD3*, Katy Milne5*, Adele Telenius, MPhil, BSc6*, Luke O`Brien7*, Celia Strong8*, Talia Goodyear8*, Shaocheng Wu2*, Katsuyoshi Takata, MD, PhD9*, Tomoko Miyata-Takata2*, Merrill Boyle, BSc10*, Susana Ben-Neriah2*, Andrew P. Weng, MD2*, Andrew Roth, PhD2*, Anca Prica, MD, MSc11*, Robert Kridel11, Brad Nelson12*, Pedro Farinha, MD, PhD3, Kerry J. Savage, MD, MSc, BSc13, David W. Scott, MBChB, PhD3 and Christian Steidl, MD, PhD6

1Princess Margaret Cancer Centre, Toronto, ON, Canada
2BC Cancer, Vancouver, Canada
3BC Cancer, Vancouver, BC, Canada
4Hematology departement, institut Curie, Paris, France
5Deeley Research Centre, BC Cancer, Victoria, BC, CAN
6Centre for Lymphoid Cancer, BC Cancer, Vancouver, BC, Canada
7BC Cancer, Centre for Lymphoid Cancer, Vancouver, BC, Canada
8Deeley Research Centre, BC Cancer, Victoria, Canada
9Division of Molecular and Cellular Pathology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
10Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada
11Division of Medical Oncology and Hematology, Princess Margaret Cancer Centre - University Health Network, Toronto, Canada
12University of Victoria, Victoria, Canada
13Division of Medical Oncology, BC Cancer, Vancouver, BC, Canada


Classic Hodgkin Lymphoma (CHL) is currently classified into histological subtypes based on morphology and antigen marker expression. However, additional biological features might help guide treatment strategies and inform prognosis. Here, we aimed to uncover disease heterogeneity and biologic subtypes of CHL based on multi-dimensional molecular profiling (Fig A) capturing somatic gene mutations, malignant cell expression phenotypes and altered tumor microenvironment (TME) architecture.


We performed exome/targeted sequencing on flow-sorted or micro-dissected HRS cells from fresh-frozen CHL biopsies (n=116) of patients treated at BC Cancer. In addition, we constructed tissue microarrays (TMA) from this cohort on which we performed GeoMx® Whole Transcriptome Assay (WTA) to obtain gene expression profiles from CD30+ HRS cells and multiplexed imaging analyses using multi-colour immunofluorescence (mIF) to delineate the spatial TME ecosystem. Cox regression analysis was performed to identify outcome differences according to molecular features in CHL patients treated with ABVD-like chemotherapy (n=104).


Mutational and copy number analyses identified known recurrent driver events including mutations and copy number changes in SOCS1, STAT6, TNFAIP3, B2M, REL, and the PDL1/PDL2 locus. These mutational findings provided a statistically powered framework to investigate the association between genomic alterations with pathological and clinical disease parameters including outcomes. Mutations affecting BCL7A, TNFAIP3, and 2p15 (REL) amplifications were strongly enriched in EBV- CHL compared to EBV+ CHL. In line with “thymic mutations” reported in grey zone lymphoma, STAT6, GNA13, ITPKB, and TBL1XR1 mutations were significantly enriched in CHL with involvement of the anterior mediastinum. ZNF217, SPEN and CD58 mutations were significantly associated with progression-free survival (PFS) (all P < .01). When investigating outcome correlation according to age group, genomic alteration in STAT6, including both single nucleotide variants and STAT6 amplification, was the most significant feature associated with unfavorable PFS in patients <45 years of age (P = .013).

Alterations in the antigen recognition system, including loss of major histocompatibility complex (MHC) is one of the most studied mechanisms of immune escape in CHL. Mutations in CSF2RB, GNA13, TNFAIP3, and STAT6 were significantly associated with MHC-I loss. Strikingly, all cases harboring CSF2RB mutations (n=23) also showed loss of MHC-I. GeoMx analysis revealed that IFN-a and IFN-g pathways were significantly upregulated in MHC-I+ HRS cells. Mutations in HIST1H1E and BCL7A were significantly enriched in cases with loss of both MHC-I and MHC-II expression.

To define mutational subtypes of CHL, we next applied non-negative matrix factorization (NMF) consensus clustering and discovered four robust subsets of tumors (clusters) using recurrent genomic events. Cluster1 (C1) is characterized by mutations in TNFAIP3, CSF2RB and GNA13, younger age, loss of MHC-I, and up-regulation of GCB and fatty acid metabolism signatures. C2 was associated with old age at initial diagnosis, EBV positivity and significant upregulation of the IFN-g pathway. C3 is characterized by REL and STAT6 amplification, and expression of a DNA repair signature. C4 includes samples from mostly younger patients with STAT6 and B2M mutations and high expression of CCL17 by HRS cells. MIF analyses further identified correlations between each mutational NMF cluster and TME composition (Fig B), and in particular specific CD4+ T cell subsets. The TME of tumors was significantly enriched by C1: FOXP3+Tregs, C2: LAG3+CD4+ cells, and C3: PD1+CD4+ cells. C4 tumors showed no positive correlation with any of these immune cell compartments. C2 tumors were also significantly associated with the abundance of CD68+ macrophages.


Our multi-dimensional profiling approach enabled us to delineate molecular profiles of HRS cells that are linked to distinct TME patterns. These linkages have implications for current pathogenesis models, molecular subtyping of CHL, and identification of cellular vulnerabilities that might be therapeutically exploitable via targeting of HRS cell phenotypes and/or immune escape mechanisms.

Disclosures: Sarkozy: AbbVie: Honoraria; BMS: Consultancy; Janssen: Consultancy; Incyte Bioscience: Consultancy, Other: Travel, Accommodations, Expenses; GSK: Consultancy; Gilead: Other: Congress fees; Roche: Other: Travel, Accommodations, Expenses, Research Funding; Prelude Therapeutics: Consultancy; Beigene: Consultancy; Lilly: Honoraria; Gilead: Other: Travel, Accommodations, Expenses; Takeda: Other: Travel, Accommodations, Expenses. Prica: Abbvie: Honoraria; Astra-Zeneca: Honoraria; Kite Gilead: Honoraria. Savage: BMS: Consultancy, Honoraria, Research Funding; Merck: Consultancy, Honoraria; Astra Zeneca: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Abbvie: Consultancy, Honoraria; Seagen: Honoraria; Roche: Research Funding. Scott: Abbvie, AstraZeneca, Incyte: Consultancy; Janssen and Roche: Research Funding. Steidl: Seattle Genetics, AbbVie, and Bayer: Consultancy; Bristol Myers Squibb, Epizyme and Trillium Therapeutics Inc.: Research Funding.

*signifies non-member of ASH