title: Flow sorting and exome sequencing reveal the oncogenome of primary Hodgkin and Reed-Sternberg cells.
abstract: Classical Hodgkin lymphoma (cHL) is characterized by sparsely distributed Hodgkin and Reed-Sternberg (HRS) cells amid reactive host background, complicating the acquisition of neoplastic DNA without extensive background contamination. We overcame this limitation by using flow-sorted HRS and intratumor T cells and optimized low-input exome sequencing of 10 patient samples to reveal alterations in genes involved in antigen presentation, chromosome integrity, transcriptional regulation, and ubiquitination. β-2-microglobulin (B2M) is the most commonly altered gene in HRS cells, with 7 of 10 cases having inactivating mutations that lead to loss of major histocompatibility complex class I (MHC-I) expression. Enforced wild-type B2M expression in a cHL cell line restored MHC-I expression. In an extended cohort of 145 patients, the absence of B2M protein in the HRS cells was associated with lower stage of disease, younger age at diagnosis, and better overall and progression-free survival. B2M-deficient cases encompassed most of the nodular sclerosis subtype cases and only a minority of mixed cellularity cases, suggesting that B2M deficiency determines the tumor microenvironment and may define a major subset of cHL that has more uniform clinical and morphologic features. In addition, we report previously unknown genetic alterations that may render selected patients sensitive to specific targeted therapies.
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY; Tri-Institutional Training Program in Computational Biology and Medicine, New York, NY;
authors.forename: Jonathan
authors.identifier: http://orcid.org/0000-0003-2613-7497
authors.initials: J
authors.lastname: Reichel
authors.affiliation: Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL;
authors.forename: Amy
authors.initials: A
authors.lastname: Chadburn
authors.affiliation: John H. Stroger Jr Hospital of Cook County, Rush University Medical Center, Ruth M. Rothstein CORE Center, Chicago, IL;
authors.forename: Paul G
authors.initials: PG
authors.lastname: Rubinstein
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY; Department of Pediatrics, Weill Cornell Medical College, New York, NY;
authors.forename: Lisa
authors.initials: L
authors.lastname: Giulino-Roth
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Wayne
authors.initials: W
authors.lastname: Tam
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Yifang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY; Botucatu School of Medicine, Sao Paulo State University, Botucatu, Brazil;
authors.forename: Rafael
authors.initials: R
authors.lastname: Gaiolla
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Kenneth
authors.initials: K
authors.lastname: Eng
authors.affiliation: Icahn School of Medicine at Mount Sinai, New York, NY;
authors.forename: Joshua
authors.initials: J
authors.lastname: Brody
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Giorgio
authors.initials: G
authors.lastname: Inghirami
authors.affiliation: Humanitas Cancer Center, Humanitas Clinical and Research Center, Rozzano (Milan), Italy; School of Medicine, University of Milan, Milan, Italy; and.
authors.forename: Carmelo
authors.initials: C
authors.lastname: Carlo-Stella
authors.affiliation: Humanitas Cancer Center, Humanitas Clinical and Research Center, Rozzano (Milan), Italy;
authors.forename: Armando
authors.initials: A
authors.lastname: Santoro
authors.affiliation: Humanitas Cancer Center, Humanitas Clinical and Research Center, Rozzano (Milan), Italy;
authors.forename: Daoud
authors.initials: D
authors.lastname: Rahal
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Jennifer
authors.initials: J
authors.lastname: Totonchy
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY; Institute for Computational Biomedicine, Weill Cornell Medical College, New York, NY.
authors.forename: Olivier
authors.initials: O
authors.lastname: Elemento
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Ethel
authors.identifier: http://orcid.org/0000-0003-3303-6299
authors.initials: E
authors.lastname: Cesarman
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY;
authors.forename: Mikhail
authors.initials: M
authors.lastname: Roshal
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D045744:Cell Line, Tumor; D002469:Cell Separation; D002648:Child; D015331:Cohort Studies; D059472:Exome / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D052138:Genes, Neoplasm; D015894:Genome, Human; D059014:High-Throughput Nucleotide Sequencing; D006689:Hodgkin Disease / Q000235:genetics* / Q000473:pathology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016539:Reed-Sternberg Cells / Q000378:metabolism* / Q000473:pathology*; D055815:Young Adult