Abstract 3638: Integrated single-cell and spatial analysis reveal the mechanism of KRASG12C inhibitor AMG510 in HSC-humanized NOG-EXL model
Authors: Wang Panpan , he Huacng , Wang Jingjing , Ni Ting , Lu Dan , Fan Qiangqiang , Shi Wenting , Gu Qingyang , Zhang Qiyao 
Published: April 2023

Background: Intra-tumor heterogeneity is one of the biggest challenges in cancer treatment today. Current single-cell RNA sequencing technologies itself could enable the exploration of gene expression heterogeneity at the single cell level, but they cannot require the morphological information within the tumor, nor the protein profiles of individual cells. Spatial transcriptomics (ST) is an in situ RNA capturing technique which preserved the spatial information of histological tissue section, and CITE-Seq is a method to preform scRNA sequencing along with acquiring protein information on a single cell level. Combining CITE-Seq and ST together can enable us to understand the roles of specific cell subpopulations and their interactions in tumor microenvironment at RNA, protein and spatial level at the same time. KRAS is the most frequently mutated oncogene in human cancer, and AMG510 is a small covalent specific KRASG12C inhibitors by blocking KRAS in its GDP-bound state, which leads to the regression of tumors and improved the anti-tumor efficacy. But the intrinsic mechanism of AMG510 in KRASG12C mutant non-small cell lung cancer (NSCLC) is not fully understood. Therefore, we develop a multi-omics method to find the mechanism of KRASG12C inhibitor AMG510 in NSCLC.

Material and Method: We established a NSCLC HSC-humanized NOG-EXL model, treated it with vehicle and AMG510, and collected 3 tumor tissues from each group. Samples were processed and cDNA library of CITE-seq were constructed based on the protocol of TotalSeq-B Human Universal Cocktail (BioLegend) and Single Cell 3’ Reagent Kits v3.1 (10x Genomics). Spatial sample preparation and cDNA library generation was carried out following the user guide of Visium FFPE Spatial Gene Expression Reagent Kits (10X Genomics). All the final libraries were sequenced by NovaSeq platform. After obtained feature-cell/spot matrix, data preprocessing and analysis were performed in R environment. Cell components in the tumor microenvironment were annotated by a cross-validated multi-approaches using CITE-seq data. And ST data were further analyzed by integrating CITE-seq data.

Results: Using a multi-omics analysis system, firstly, we constructed a single-cell transcriptomic atlas of NSCLC tumor microenvironment; and then explored the characterization of cell-type infiltration. Furthermore, CITE-seq and ST datasets were integrated to assess the spatial organization of infiltrated immune cells and tumor cells. We found how AMG510 affected tumor microenvironment and enhanced the checkpoint pathways. The results in return proved the reliability of the multi-omics system, which enables us to explore how drugs treat cancer by affecting its microenvironment in multiple levels.

Summary: We established a multi-omics analysis system and explored the mechanism of the KRASG12C inhibitor AMG510 against the NSCLC.

Citation Format: Panpan Wang, Huacng he, Jingjing Wang, Ting Ni, Dan Lu, Qiangqiang Fan, Wenting Shi, Qingyang Gu, Qiyao Zhang. Integrated single-cell and spatial analysis reveal the mechanism of KRASG12C inhibitor AMG510 in HSC-humanized NOG-EXL model. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3638.