Single-Cell RNA Sequencing Identifies ALCAM and Gpnmb as Therapeutic Targets for ADC Development∗bull;∗bull;∗bull; in Squamous Cell Carcinoma of the Lung

Document Type

Conference Proceeding

Publication Date

10-1-2025

Publication Title

J Thorac Oncol

Keywords

activated leukocyte cell adhesion molecule, antibody drug conjugate, breast cancer resistance protein, camptothecin, clathrin, DNA topoisomerase, emtansine, glembatumumab vedotin, maytansine derivative, monoclonal antibody, praluzatamab ravtansine, sialic acid, vedotin, CD8+ T lymphocyte, cell surface, conference abstract, controlled study, endocytosis, epithelium cell, fibroblast, human, human cell, human tissue, lung adenocarcinoma, lung alveolus cell, lung carcinoma, microtubule, mitochondrial gene, non small cell lung cancer, principal component analysis, single cell RNA seq, squamous cell carcinoma, tumor microenvironment

Abstract

Introduction: Antibody-drug conjugates (ADCs) combine the specificity of monoclonal antibodies with the cytotoxic potency of chemotherapeutic agents and have emerged as a promising treatment strategy in oncology. In non-small cell lung cancer (NSCLC), several ADCs have been investigated and showed better responses in lung adenocarcinoma (LUAD) compared to squamous cell carcinoma (LUSC). In this analysis, we leverage single-cell RNA sequencing to identify optimal targets and payloads for ADC development in LUSC. Methods: The raw cell expression matrix, cellular annotations, and metadata for GSE131907, GSE148071, and GSE162498 were downloaded from TISCH2.0. Cells with less than 500 genes, 1000 counts, or more than 10 percent mitochondrial genes were excluded. A total of 361,000 cells from 97 patients were utilized. Preprocessing included merging the datasets, normalizing, scaling, and running principal component analysis. Integration of the datasets was then carried out using harmony, adjusting for batch effects. Differential expression of ADC targets between cell types and tissue subtypes was performed using the FindMarkers function in Seurat and a limma-based pseudobulk approach. Correlation analysis between ADC targets and sensitivity/resistance markers to topoisomerase 1 and microtubule inhibitors was conducted using the CSCORE algorithm. Results: Transmembrane glycoprotein NMB (GPNMB) was significantly overexpressed in LUSC malignant cells versus normal epithelial cells (log2FC = 1.10, P = 0.012), while Activated leukocyte cell adhesion molecule (ALCAM) did not reach statistical significance. Both targets were also elevated in multiple cell types in the LUSC tumor microenvironment as compared to LUAD, including fibroblasts, macrophages, alveolar cells, and CD8+ T cells (p < 0.0001). In LUSC malignant epithelial cells, ALCAM negatively correlated with ABCC2 (r = -0.41, P < 0.0001) and ABCB1 (r = -0.17, P < 0.0001), two efflux transporters linked to resistance to maytansinoids and emtansine. ALCAM also positively correlated with CLTC (r = 0.96, P < 0.0001), which mediates clathrin-dependent ADC internalization. In contrast, GPNMB showed strong positive correlations with ABCG2, ABCC2, ABCB1, and SLC47A2 (all p <0.0001), which confer resistance to MMAE and camptothecins via drug efflux. Lastly, GPNMB (r = -1, p < 0.0001) and ALCAM (r = -0.79, p < 0.0001) were negatively corre- lated with NANS expression which can lead to increased ADC internalization and sensitivity by depleting sialic acid from the cell surface. Conclusions: Our analysis highlights ALCAM and GPNMB as potential ADC targets in LUSC, with broad expression across malignant and stromal compartments. ALCAM's expression profile suggests suscep- tibility to maytansinoid-based ADCs such as Praluzatamab Ravtansine, due to its association with endocytosis and absence of efflux resis- tance signals. GPNMB, while a promising target, is associated with expression of multiple efflux transporters, raising concerns about resistance to ADCs like Glembatumumab Vedotin. These findings support a tailored approach to ADC design based on tumor subtype and resistance landscape.

Volume

20

Issue

10

First Page

S376

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