Transcriptional and functional analyses of neoantigen-specific CD4 T cells during a profound response to anti-PD-L1 in metastatic Merkel cell carcinoma.
Transcriptional and functional analyses of neoantigen-specific CD4 T cells during a profound response to anti-PD-L1 in metastatic Merkel cell carcinoma.
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DOI：
10.1136/jitc-2022-005328
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发表时间：
2022-09
影响因子：
10.9
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中科院分区：
文献类型：
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默克尔细胞癌（MCC）通常对PD-1通路阻断有反应，与肿瘤病毒状态无关（约80%的病例由默克尔细胞多瘤病毒（MCPyV）驱动）。先前的研究已经描述了肿瘤特异性T细胞对MCPyV的反应，通常是CD8，但对T细胞对紫外线诱导的新抗原的反应知之甚少。一位50多岁的病毒阴性（VN） MCC患者在短暂的化疗初期反应后出现了大面积的肝转移。她接受了抗pd - l1 （avelumab）治疗，并在4周内部分缓解。采用全外显子组测序（WES）确定潜在的新抗原肽。通过干扰素γ (IFNγ) ELISpot、流式细胞术和单细胞RNA测序评估外周血新抗原T细胞反应的特征。肿瘤驻留T细胞采用多重免疫组织化学表征。WES鉴定出1027个肿瘤特异性体细胞突变，与已发表的vn - mcc平均1121个相似。结合截止值≤100 nM的肽预测结果合成了77个用于T细胞检测的肽。虽然多肽是基于I类hla预测的，但我们发现循环CD4 T细胞靶向77种新抗原中的5种。相比之下，没有检测到新抗原特异性CD8 T细胞反应。在抗pd - l1治疗前，血液中检测不到新抗原特异性CD4 T细胞，但在开始治疗后不久就很容易检测到。T细胞在新抗原（突变）肽刺激下产生强大的IFNγ，而在正常（野生型）肽刺激下则不产生。单细胞RNAseq显示新抗原反应性T细胞表达th1相关转录因子（T-bet）和相关细胞因子。这些CD4 T细胞没有明显的细胞毒性或非th1标记物。在预处理肿瘤中，常驻CD4 T细胞也呈th1偏斜并表达T-bet。我们鉴定并表征了针对多种新抗原的肿瘤特异性th1倾斜CD4 T细胞，该患者对抗pd - l1治疗产生了深刻而持久的部分反应。据我们所知，这是首个关于MCC中新抗原特异性T细胞反应的报道。虽然识别病毒肿瘤抗原的CD4和CD8 T细胞在病毒阳性的MCC中经常被检测到，但在该患者中仅检测到识别新抗原的CD4 T细胞。这些发现提示CD4 T细胞在抗pd -(L)1治疗应答中发挥重要作用。
Merkel cell carcinoma (MCC) often responds to PD-1 pathway blockade, regardless of tumor-viral status (~80% of cases driven by the Merkel cell polyomavirus (MCPyV)). Prior studies have characterized tumor-specific T cell responses to MCPyV, which have typically been CD8, but little is known about the T cell response to UV-induced neoantigens. A patient in her mid-50s with virus-negative (VN) MCC developed large liver metastases after a brief initial response to chemotherapy. She received anti-PD-L1 (avelumab) and had a partial response within 4 weeks. Whole exome sequencing (WES) was performed to determine potential neoantigen peptides. Characterization of peripheral blood neoantigen T cell responses was evaluated via interferon-gamma (IFNγ) ELISpot, flow cytometry and single-cell RNA sequencing. Tumor-resident T cells were characterized by multiplexed immunohistochemistry. WES identified 1027 tumor-specific somatic mutations, similar to the published average of 1121 for VN-MCCs. Peptide prediction with a binding cut-off of ≤100 nM resulted in 77 peptides that were synthesized for T cell assays. Although peptides were predicted based on class I HLAs, we identified circulating CD4 T cells targeting 5 of 77 neoantigens. In contrast, no neoantigen-specific CD8 T cell responses were detected. Neoantigen-specific CD4 T cells were undetectable in blood before anti-PD-L1 therapy but became readily detectible shortly after starting therapy. T cells produced robust IFNγ when stimulated by neoantigen (mutant) peptides but not by the normal (wild-type) peptides. Single cell RNAseq showed neoantigen-reactive T cells expressed the Th1-associated transcription factor (T-bet) and associated cytokines. These CD4 T cells did not significantly exhibit cytotoxicity or non-Th1 markers. Within the pretreatment tumor, resident CD4 T cells were also Th1-skewed and expressed T-bet. We identified and characterized tumor-specific Th1-skewed CD4 T cells targeting multiple neoantigens in a patient who experienced a profound and durable partial response to anti-PD-L1 therapy. To our knowledge, this is the first report of neoantigen-specific T cell responses in MCC. Although CD4 and CD8 T cells recognizing viral tumor antigens are often detectible in virus-positive MCC, only CD4 T cells recognizing neoantigens were detected in this patient. These findings suggest that CD4 T cells can play an important role in the response to anti-PD-(L)1 therapy.
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DOI：
10.1084/jem.188.12.2357
发表时间：
1998-12-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Hung K;Hayashi R;Lafond-Walker A;Lowenstein C;Pardoll D;Levitsky H
通讯作者：
Levitsky H
影响因子：
8.4
作者：
Eisenhauer, E. A.;Therasse, P.;Verweij, J.
通讯作者：
Verweij, J.
影响因子：
15.9
作者：
Lee, Patrick C.;Klaeger, Susan;Le, Phuong M.;Korthauer, Keegan;Cheng, Jingwei;Ananthapadmanabhan, Varsha;Frost, Thomas C.;Stevens, Jonathan D.;Wong, Alan Y. L.;Iorgulescu, J. Bryan;Tarren, Anna Y.;Chea, Vipheaviny A.;Carulli, Isabel P.;Lemvigh, Camilla K.;Pedersen, Christina B.;Gartin, Ashley K.;Sarkizova, Siranush;Wright, Kyle T.;Li, Letitia W.;Nomburg, Jason;Li, Shuqiang;Huang, Teddy;Liu, Xiaoxi;Pomerance, Lucas;Doherty, Laura M.;Apffel, Annie M.;Wallace, Luke J.;Rachimi, Suzanna;Felt, Kristen D.;Wolff, Jacquelyn O.;Witten, Elizabeth;Zhang, Wandi;Neuberg, Donna;Lane, William J.;Zhang, Guanglan;Olsen, Lars R.;Thakuria, Manisha;Rodig, Scott J.;Clauser, Karl R.;Starrett, Gabriel J.;Doench, John G.;Buhrlage, Sara J.;Carr, Steven A.;DeCaprio, James A.;Wu, Catherine J.;Keskin, Derin B.
通讯作者：
Keskin, Derin B.
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
24.8
作者：
Ahmadzadeh, Mojgan;Pasetto, Anna;Rosenberg, Steven A.
通讯作者：
Rosenberg, Steven A.