title: Deep Sequencing of T-cell Receptor DNA as a Biomarker of Clonally Expanded TILs in Breast Cancer after Immunotherapy.
abstract: In early-stage breast cancer, the degree of tumor-infiltrating lymphocytes (TIL) predicts response to chemotherapy and overall survival. Combination immunotherapy with immune checkpoint antibody plus tumor cryoablation can induce lymphocytic infiltrates and improve survival in mice. We used T-cell receptor (TCR) DNA sequencing to evaluate both the effect of cryoimmunotherapy in humans and the feasibility of TCR sequencing in early-stage breast cancer. In a pilot clinical trial, 18 women with early-stage breast cancer were treated preoperatively with cryoablation, single-dose anti-CTLA-4 (ipilimumab), or cryoablation + ipilimumab. TCRs within serially collected peripheral blood and tumor tissue were sequenced. In baseline tumor tissues, T-cell density as measured by TCR sequencing correlated with TIL scores obtained by hematoxylin and eosin (H&E) staining. However, tumors with little or no lymphocytes by H&E contained up to 3.6 × 106TCR DNA sequences, highlighting the sensitivity of the ImmunoSEQ platform. In this dataset, ipilimumab increased intratumoral T-cell density over time, whereas cryoablation ± ipilimumab diversified and remodeled the intratumoral T-cell clonal repertoire. Compared with monotherapy, cryoablation plus ipilimumab was associated with numerically greater numbers of peripheral blood and intratumoral T-cell clones expanding robustly following therapy. In conclusion, TCR sequencing correlates with H&E lymphocyte scoring and provides additional information on clonal diversity. These findings support further study of the use of TCR sequencing as a biomarker for T-cell responses to therapy and for the study of cryoimmunotherapy in early-stage breast cancer. Cancer Immunol Res; 4(10); 835-44. ©2016 AACR.
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York. Providence Portland/Robert W. Franz Cancer Research Center and Earl A. Chiles Research Institute, Portland, Oregon.
authors: David B
authors: DB
authors: Page
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Jianda
authors: J
authors: Yuan
authors: Weill Cornell Medical College, New York, New York. david.page2@providence.org.
authors: David
authors: D
authors: Redmond
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Y Hanna
authors: YH
authors: Wen
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Jeremy C
authors: JC
authors: Durack
authors: Adaptive Biotechnologies, Seattle, Washington.
authors: Ryan
authors: R
authors: Emerson
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Stephen
authors: S
authors: Solomon
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Zhiwan
authors: Z
authors: Dong
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Phillip
authors: P
authors: Wong
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Christopher
authors: C
authors: Comstock
authors: The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors: Adi
authors: A
authors: Diab
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Janice
authors: J
authors: Sung
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Majid
authors: M
authors: Maybody
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Elizabeth
authors: E
authors: Morris
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Edi
authors: E
authors: Brogi
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Monica
authors: M
authors: Morrow
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Virgilio
authors: V
authors: Sacchini
authors: Weill Cornell Medical College, New York, New York.
authors: Olivier
authors: O
authors: Elemento
authors: Fred Hutchinson Cancer Center, Seattle, Washington.
authors: Harlan
authors: H
authors: Robins
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York.
authors: Sujata
authors: S
authors: Patil
authors: The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors: James P
authors: JP
authors: Allison
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York. Weill Cornell Medical College, New York, New York.
authors: Jedd D
authors: JD
authors: Wolchok
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York. Weill Cornell Medical College, New York, New York.
authors: Clifford
authors: C
authors: Hudis
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York. Weill Cornell Medical College, New York, New York.
authors: Larry
authors: L
authors: Norton
authors: Memorial Sloan Kettering Cancer Center (MSKCC), New York, New York. Weill Cornell Medical College, New York, New York.
authors: Heather L
authors: HL
authors: McArthur
mesh_terms: D000074322:Antineoplastic Agents, Immunological / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000235:genetics*; D001706:Biopsy; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D003131:Combined Modality Therapy; D003452:Cryosurgery; D004273:DNA, Neoplasm / Q000235:genetics; D005240:Feasibility Studies; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D000074324:Ipilimumab / Q000627:therapeutic use; D017254:Leukemic Infiltration; D018655:Lymphocyte Count; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008408:Mastectomy / Q000379:methods; D009367:Neoplasm Staging; D010865:Pilot Projects; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D000074322:Antineoplastic Agents, Immunological; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D000074324:Ipilimumab; D016693:Receptors, Antigen, T-Cell, alpha-beta