GITR and TIGIT immunotherapy provokes divergent multicellular responses in the tumor microenvironment of gastrointestinal cancers.
GITR and TIGIT immunotherapy provokes divergent multicellular responses in the tumor microenvironment of gastrointestinal cancers.
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DOI：
10.1186/s13073-023-01259-3
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发表时间：
2023-11-26
期刊：
影响因子：
12.3
通讯作者：
中科院分区：
文献类型：
--
作者：
了解新型免疫治疗剂的机制效应对于改善其成功的临床转化至关重要。这些效应需要在临床前模型中进行研究，这些模型维持了患者肿瘤中发现的异质性肿瘤微环境（TME）和功能失调的细胞状态。我们研究了在患者来源的离体系统中靶向共刺激分子GITR和共抑制免疫检查点TIGIT的免疫治疗扰动，该系统将TME维持在其接近天然状态。利用单细胞基因组学，我们确定了细胞类型特异性转录重编程，以响应免疫治疗扰动。我们从新鲜的胃癌和结肠癌手术切除物中产生了离体肿瘤切片培养物，并用GITR激动剂或TIGIT拮抗剂抗体对其进行处理。我们将配对的单细胞RNA和TCR测序应用于原始手术切除、对照和经处理的离体肿瘤切片培养物。我们还使用多重免疫荧光证实了靶点表达，并使用RNA原位杂交验证了我们的发现。我们证实，肿瘤切片培养物保持了原始手术切除的细胞类型、转录细胞状态和比例。GITR激动剂仅限于增加细胞毒性CD 8 T细胞中的效应基因表达。功能失调的耗尽的CD 8 T细胞对GITR激动剂没有应答。相比之下，TIGIT拮抗剂增加TCR信号传导并活化细胞毒性和功能失调的CD 8 T细胞。这包括对应于具有指示潜在肿瘤抗原反应性的特征的TCR克隆型的细胞。TIGIT拮抗剂还活化T滤泡辅助细胞样细胞和树突细胞，并减少调节性T细胞中的免疫抑制标志物。我们鉴定了GITR和TIGIT免疫疗法在患者的TME中的新的细胞作用机制。与产生有限转录应答的GITR激动剂不同，TIGIT拮抗剂协调涉及CD 8 T细胞、T滤泡辅助细胞样细胞、树突细胞和调节性T细胞的多细胞应答。我们将单细胞基因组学与临床前模型相结合的实验策略可以成功地识别新型免疫治疗剂的作用机制。了解反应或抗性的细胞和转录机制将有助于优先考虑靶点及其临床翻译。在线版本包含补充材料，可通过10.1186/s13073-023-01259-3获得。
Understanding the mechanistic effects of novel immunotherapy agents is critical to improving their successful clinical translation. These effects need to be studied in preclinical models that maintain the heterogenous tumor microenvironment (TME) and dysfunctional cell states found in a patient’s tumor. We investigated immunotherapy perturbations targeting co-stimulatory molecule GITR and co-inhibitory immune checkpoint TIGIT in a patient-derived ex vivo system that maintains the TME in its near-native state. Leveraging single-cell genomics, we identified cell type-specific transcriptional reprogramming in response to immunotherapy perturbations. We generated ex vivo tumor slice cultures from fresh surgical resections of gastric and colon cancer and treated them with GITR agonist or TIGIT antagonist antibodies. We applied paired single-cell RNA and TCR sequencing to the original surgical resections, control, and treated ex vivo tumor slice cultures. We additionally confirmed target expression using multiplex immunofluorescence and validated our findings with RNA in situ hybridization. We confirmed that tumor slice cultures maintained the cell types, transcriptional cell states and proportions of the original surgical resection. The GITR agonist was limited to increasing effector gene expression only in cytotoxic CD8 T cells. Dysfunctional exhausted CD8 T cells did not respond to GITR agonist. In contrast, the TIGIT antagonist increased TCR signaling and activated both cytotoxic and dysfunctional CD8 T cells. This included cells corresponding to TCR clonotypes with features indicative of potential tumor antigen reactivity. The TIGIT antagonist also activated T follicular helper-like cells and dendritic cells, and reduced markers of immunosuppression in regulatory T cells. We identified novel cellular mechanisms of action of GITR and TIGIT immunotherapy in the patients’ TME. Unlike the GITR agonist that generated a limited transcriptional response, TIGIT antagonist orchestrated a multicellular response involving CD8 T cells, T follicular helper-like cells, dendritic cells, and regulatory T cells. Our experimental strategy combining single-cell genomics with preclinical models can successfully identify mechanisms of action of novel immunotherapy agents. Understanding the cellular and transcriptional mechanisms of response or resistance will aid in prioritization of targets and their clinical translation. The online version contains supplementary material available at 10.1186/s13073-023-01259-3.
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影响因子：
4.6
作者：
Bankhead P;Loughrey MB;Fernández JA;Dombrowski Y;McArt DG;Dunne PD;McQuaid S;Gray RT;Murray LJ;Coleman HG;James JA;Salto-Tellez M;Hamilton PW
通讯作者：
Hamilton PW
影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
30.5
作者：
Galletti G;De Simone G;Mazza EMC;Puccio S;Mezzanotte C;Bi TM;Davydov AN;Metsger M;Scamardella E;Alvisi G;De Paoli F;Zanon V;Scarpa A;Camisa B;Colombo FS;Anselmo A;Peano C;Polletti S;Mavilio D;Gattinoni L;Boi SK;Youngblood BA;Jones RE;Baird DM;Gostick E;Llewellyn-Lacey S;Ladell K;Price DA;Chudakov DM;Newell EW;Casucci M;Lugli E
通讯作者：
Lugli E
用于抗癌药物开发的器官型组织切片培养的最新进展。
影响因子：
9.2
作者：
He L;Deng C
通讯作者：
Deng C
影响因子：
5.6
作者：
den Braanker H;van Stigt AC;Kok MR;Lubberts E;Bisoendial RJ
通讯作者：
Bisoendial RJ