PMID- 38395378 OWN - NLM STAT- MEDLINE DCOM- 20240409 LR - 20240409 IS - 1872-7980 (Electronic) IS - 0304-3835 (Linking) VI - 588 DP - 2024 Apr 28 TI - Antigen/HLA-agnostic strategies for Characterizing Tumor-responsive T cell receptors in PDAC patients via single-cell sequencing and autologous organoid application. PG - 216741 LID - S0304-3835(24)00134-4 [pii] LID - 10.1016/j.canlet.2024.216741 [doi] AB - Characterization of tumor-responsive T cell receptors (TCRs) is a critical step in personalized TCR-T cell therapy, and remains challenging for pancreatic ductal adenocarcinoma (PDAC). Here we report a proof-of-concept study to identify and validate antitumor TCRs in two representative PDAC patients using ultradeep single-cell TCR/RNA sequencing and autologous organoids, and reveal the phenotypic dynamics of TCR repertoire in different T cell expansions from the same patient. We first performed comparative sequencing on freshly harvested peripheral blood mononuclear cells (PBMCs) and uncultured tumor infiltrating lymphocytes (TILs), followed by reactivity tests of TIL-enriched TCRs with autologous organoids, in which two tumor-responsive TCRs were successfully characterized and the corresponding TILs were mostly tissue-resident memory-like T cells, and partially expressed both naive and exhausted T cell markers. For the PDAC patient without high-quality TILs, PBMCs were cultured with neoantigen peptide (KRAS(G12D)), organoids, or anti-CD3 antibody in presence, and experienced extensive clonal expansions within ten days. All derived PBMCs were sequenced in parallel (>82,000 cells), and TCRs enriched in both peptide- and organoid-experienced, but not anti-CD3-treated CD8 T cells, were assessed for their reactivity to antigen-presenting cells (APCs) and organoids, in which three neoantigen-reactive TCRs were identified as tumor-responsive, and the corresponding T cells were characterized by mixed transcriptional signatures including but not limited to typical exhausted T cell markers. Together, our study revealed that the combination of ultradeep single-cell sequencing and organoid techniques enabled rapid characterization of tumor-responsive TCRs for developing practical personalized TCR-T therapy in an antigen/human leukocyte antigen (HLA)-agnostic manner. CI - Copyright (c) 2024 Elsevier B.V. All rights reserved. FAU - Wang, Xu AU - Wang X AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China; Cancer Institute, Shanghai Key Laboratory of Radiation Oncology, Cancer Research Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China. FAU - Dai, Zhengjie AU - Dai Z AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Lin, Xuan AU - Lin X AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Zou, Xuan AU - Zou X AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Wang, Ruijie AU - Wang R AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Tasiheng, Yesboli AU - Tasiheng Y AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Yan, Yu AU - Yan Y AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Ma, Mingjian AU - Ma M AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Chen, Yusheng AU - Chen Y AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. FAU - Cheng, He AU - Cheng H AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. Electronic address: chenghe@fudanpci.org. FAU - Liu, Chen AU - Liu C AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. Electronic address: liuchen@fudanpci.org. FAU - Yu, Xianjun AU - Yu X AD - Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Precise Diagnosis and Treatment of Pancreatic Cancer, Shanghai Pancreatic Cancer Institute, Shanghai, China; Pancreatic Cancer Institute, Fudan University, Shanghai, China. Electronic address: yuxianjun@fudanpci.org. LA - eng PT - Journal Article DEP - 20240222 PL - Ireland TA - Cancer Lett JT - Cancer letters JID - 7600053 RN - 0 (Receptors, Antigen, T-Cell) RN - 0 (Antigens, Neoplasm) RN - 0 (HLA Antigens) RN - 0 (CD3 Complex) RN - 0 (Histocompatibility Antigens Class II) RN - 0 (Peptides) SB - IM MH - Humans MH - *Receptors, Antigen, T-Cell/genetics MH - Lymphocytes, Tumor-Infiltrating MH - CD8-Positive T-Lymphocytes MH - Antigens, Neoplasm/genetics MH - HLA Antigens MH - *Pancreatic Neoplasms/genetics/therapy MH - CD3 Complex MH - Histocompatibility Antigens Class II MH - Peptides MH - Organoids OTO - NOTNLM OT - Neoantigen OT - Organoid OT - PDAC OT - Single-cell sequencing OT - TCR-T OT - TILs COIS- Declaration of competing interest The authors declared no conflict of interest. EDAT- 2024/02/24 11:42 MHDA- 2024/04/09 06:45 CRDT- 2024/02/23 19:19 PHST- 2023/11/08 00:00 [received] PHST- 2024/02/05 00:00 [revised] PHST- 2024/02/16 00:00 [accepted] PHST- 2024/04/09 06:45 [medline] PHST- 2024/02/24 11:42 [pubmed] PHST- 2024/02/23 19:19 [entrez] AID - S0304-3835(24)00134-4 [pii] AID - 10.1016/j.canlet.2024.216741 [doi] PST - ppublish SO - Cancer Lett. 2024 Apr 28;588:216741. doi: 10.1016/j.canlet.2024.216741. Epub 2024 Feb 22.