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4361 Neuronal-Glial and Immune Microenvironment Signatures Show Prognostic Effects in Primary CNS Lymphoma

Program: Oral and Poster Abstracts
Session: 621. Lymphomas: Translational – Molecular and Genetic: Poster III
Hematology Disease Topics & Pathways:
Research, Adult, Translational Research, Lymphomas, Non-Hodgkin lymphoma, Elderly, Clinical Research, B Cell lymphoma, Genomics, Diseases, Real-world evidence, Aggressive lymphoma, Immunology, Lymphoid Malignancies, Biological Processes, Technology and Procedures, Molecular biology, Profiling, Study Population, Human, Pathogenesis, Imaging, Machine learning, Omics technologies, Pathology
Monday, December 9, 2024, 6:00 PM-8:00 PM

Zijun Y. Xu-Monette, PhD1, Xiaoxiao Wang2*, Govind Bhagat, MD3, Alexandar Tzankov4*, Qingyan Au5*, Carlo Visco, MD6,7*, Harry Nunns8*, Alessandro La Ferlita, Ph.D9*, Lalit Sehgal, PhD10, April Chiu, M.D.11*, Heounjeong Go12*, Anamarija M. Perry, MD13, Huilan Rao2*, Mina Xu, MD14*, Qiongli Zhai15*, Yafei Wang15*, Shi Wang16*, Wenbin Qian17*, Maurilio Ponzoni, MD18,19, Wen Shuai1*, Bangchen Wang, MD1*, Sergio Pina-Oviedo, MD1*, Luis Carrillo, MD1*, Kelly Au20*, Santiago Montes-Moreno Sr., MD, PhD21*, Karen Dybkaer, PhD22*, Zenggang Pan, MD, PhD23*, William Choi, MBBS, MRCP24, Michael Møller25*, Luigi Racioppi, MD, PhD20*, Xiaohui Zhang, MD, PhD26*, Jinming Song, MD, PhD27*, Shanxiang Zhang, MD, PhD28*, Youli Zu, MD, PhD29*, Joo Y. Song, MD30, Eric D. Hsi, MD31, Fenghuang Zhan, MD32, Benjamin M. Parsons, DO33, Aliyah R. Sohani, MD34*, Weina Chen, MD, PhD35*, Fabio Iwamoto36*, Jeremy S. Abramson, MD37, Bing Xu38*, Yong Li, PhD39 and Ken H. Young, MD, PhD40

1Department of Pathology, Duke University Medical Center, Durham, NC
2Sun Yat-Sen University Cancer Center, Guangzhou, China
3Department of Pathology & Cell Biology, Columbia University, New York, NY
4Medical University Basel, Basel, Switzerland
5NeoGenomics Laboratories, Aliso Viejo, CA
6Department of Engineering for Innovation Medicine/Section of Hematology, University of Verona, Verona, Italy
7Hematology and Bone Marrow Transplant Unit, Department of Engineering for Innovative Medicine, University of Verona, Verona, Italy
8Neogenomics Laboratories, Aliso Viejo, CA
9Division of Hematology, College of Medicine, The Ohio State University, Columbus, OH
10James Comprehensive Cancer Center, The Ohio State University, Columbus, OH
11Division of Hematopathology, Mayo Clinic, Rochester, MN
12Asan Medical Center Children's Hospital, Seoul, KOR
13Department of Pathology, University of Michigan, Ann Arbor, MI
14Yale University, New Haven, CT
15Tianjin Medical University Cancer Institute and Hospital, Tianjin, China
16National University Hospital, Singapore, Singapore, Singapore
17The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
18Pathology Unit, San Raffaele H. Scientific Institute, Milano, Italy
19Pathology Unit, IRCCS San Raffaele Scientific Institute and Università Vita-Salute San Raffaele, Milan, Italy
20Duke University, Durham, NC
21IDIVAL/Valdecilla, on behalf of GELTAMO., Santander, Spain
22Department of Haematology, Aalborg University Hospital, Aalborg, Denmark
23Department of Pathology, University of Colorado Anschutz Medical Campus, Aurora, CO
24Tuen Mun Hospital, Happy Valley, NA, HKG
25Odense University Hospital, Odense, DNK
26Department of Pathology, H. Lee Moffitt Cancer Center, Tampa, FL
27Department of Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL
28Department of Pathology & Laboratory Medicine, Indiana University School of Medicine, Indianapolis, IN
29Houston Methodist Hospital, Houston, TX
30Department of Hematology and Hematopoietic Cell Transplantation, City of Hope National Medical Center, Duarte, CA
31Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN
32Myeloma Center, Winthrop P. Rockefeller Cancer Institute, University of Arkansas for Medical Sciences, Little Rock, AR
33Gundersen Lutheran Health System, La Crosse, WI
34Department of Pathology, Maasachusetts General Hospital, Boston, MA
35Department of Pathology, UT Southwestern Medical Center, Dallas, TX
36Columbia University Irving Medical Center, New York
37Massachusetts General Hospital Cancer Center, Boston, MA
38The First Affiliated Hospital of Xiamen University and Institute of Hematology, Xiamen University, Xiamen, China
39Department of Medicine, Baylor College of Medcine, Houston, TX
40Duke Cancer Institute, Duke University Medical Center, Durham, NC

Introduction: Primary diffuse large B-cell lymphoma (DLBCL) of the central nervous system (CNS), traditionally referred to as primary CNS lymphoma (PCNSL), is a rare type of primary large B-cell lymphoma of immune-privileged sites with significantly poorer prognosis than systemic DLBCL. Genetic profiles of PCNSL were widely studied, whereas the tumor microenvironment has not been well characterized.

Patients and Methods: A series of molecular analyses were performed using DLBCL FFPE tissues collected by the International Lymphoma Consortium. First, multiplex fluorescent immunohistochemistry (mfIHC) using antibodies for CD20, PAX5, CD3, CD4, CD8, FOXP3, CD45RO, CD68, CD56, PD-1, PD-L1, PD-L2, and CTLA-4 was performed on PCNSL tissues from 121 immunocompetent patients and de novo systemic DLBCL tissues from 405 patients. Digital quantification results for 405 patients with EBV-negative systemic DLBCL or primary testicular (PT) DLBCL have been reported previously. The preliminary mfIHC results of PCNSL will be presented. Second, mfIHC for multiple B cell biomarkers, CD3, CD4, CD8, CD68, CD163, CD11c, and CD31 were performed for 10 PCNSL validation cases, 24 PT-DLBCL and 451 systemic DLBCL tissues (unpublished). Third, RNA samples were extracted and sequenced for the targeted 1408 cancer-associated genes, successfully in 142 PCNSL cases, including 102 cases evaluated by either of these two mfIHC panels and additional 40 PCNSL cases,

Results: First, in the first large mfIHC cohort, PCNSLs compared with de novo systemic nodal or extranodal DLBCL had significantly higher mean/median CD56+ cell densities whereas lower T cell densities (consistent with the immune privilege), as well as lower CD45RO+, PD-1+, and FOXP3+ percentages in T cells whereas higher CD4¯CD8¯ double-negative percentages in T cells. Prognostic analysis in the PCNSL cohort showed that high CD56+ cell densities and high CD45RO+CD8+ T cell densities were associated with significantly poorer and better patient survival, respectively, which are opposite to their prognostic effects in systemic DLBCL as previously reported. Twelve patients had very high PD-L1 expression in B cells and significantly poorer survival in the PCNSL cohort. Second, as validation, the 10 PCNSL cases in the second mfIHC study showed significantly less T cells than systemic DLBCL and PT-DLBCL cases, and low CD8 T cell densities and having low percentage of CD8 T cells adjacent to tumor cells were associated with significantly poorer survival. Immunofluorescence landscape and spatial analysis revealed significant differences in immune privilege between PCNSL and PT-DLBCL. Third, gene expression profiling analysis by targeted RNA-seq identified 282 significantly differentially expressed genes between PCNSL and systemic DLBCL, referred to as the PCNSL signature. Unsupervised clustering analysis divided the PCNSL cohort into two clusters, which had no significant difference in MYD88 mutation frequencies analyzed at the mRNA level. Importantly, the cluster with higher expression of many neural glial genes in the PCNSL signature had significantly poorer survival, which remained significant in multivariate analysis adjusting for clinical factors and/or treatments, two PCNSL subcohorts (as training and validation cohorts), and methotrexate-treated patients. NCAM1 (Neural Cell Adhesion Molecule 1, encoding CD56) was among the upregulated PCNSL signature genes, explaining the poorer prognosis associated with higher CD56+ cell densities in PCNSL by the mfIHC analysis.

Summary: Our studies represent the largest immunophenotypic and gene expression profiling studies currently in PCNSL, a rare type of primary DLBCL of immune-privileged sites. Multiplex immunofluorescence revealed the immune landscape in PCNSL in comparison with systemic and testicular DLBCL, and identified prognostic immune biomarkers. Targeted RNA-seq identified a PCNSL gene signature enriched with neuronal-glial genes and showing significant prognostic effect in PCNSL. These novel findings in the PCNSL microenvironment warrant further studies.

Disclosures: Au: NeoGenomics Laboratories, Aliso Viejo, California, USA: Current Employment. Nunns: NeoGenomics Laboratories, Aliso Viejo, California, USA: Current Employment. Chiu: AstraZeneca: Current Employment. Xu: Treeline Biosciences: Consultancy; Evans, Haigh & Arndt LLP: Other: Paid expert testimon. Montes-Moreno: Roche: Speakers Bureau; Janssen: Speakers Bureau; Recordati: Speakers Bureau. Hsi: Abcon Therapeutics: Current holder of stock options in a privately-held company; Eli Lilly: Other: sponsored research. Abramson: Cellectis: Research Funding; Bristol Myers Squibb: Consultancy, Research Funding; Mustang Bio: Research Funding; Merck: Research Funding; Seagen Inc.: Research Funding; Interius BioTh: Consultancy; Incyte Corporation: Consultancy; Genmab US Inc: Consultancy; Genentech, a member of the Roche Group: Consultancy; AstraZeneca Pharmaceuticals LP: Consultancy; BeiGene Ltd: Consultancy; Caribou Biosciences Inc: Consultancy; Century Therapeutics: Consultancy; Epizyme Inc: Consultancy; AbbVie Inc: Consultancy; Foresight Diagnostics: Consultancy. Young: Flagship Biosciences: Consultancy; Arima Genomics: Consultancy.

*signifies non-member of ASH