Abstract 5386: Increased sprouting angiogenesis in a microphysiological model of cisplatin-resistant recurrent lung cancers

Abstract

Abstract Non-small cell lung cancer (NSCLC) constitutes 85% of total lung cancer cases and first-line therapies still often employ the platinum-based agents. Cisplatin therapy for NSCLC often leads to relapse of aggressive tumors with high metastatic potential after a period of dormancy. We hypothesize that cisplatin-resistant tumor cell phenotypes will increase features of aggressivity including sprouting angiogenesis in microphysiological models of NSCLC. First, we engineered various TME configurations using A549 cell spheroids and cancer associated fibroblasts (CAF) to establish a baseline of tumor cell proliferation, ECM deposition, and expression of proangiogenic and proinflammatory genes. Next, we generated cisplatin-resistant (Cis-R) A549 sub-lines to test the hypothesis that 3D tumor models engineered using these drug-resistant cells will capture hallmarks of highly aggressive recurrent tumors such as increased expression of progression-associated genes, increased induction of local stromal reactions and more profound angiogenesis. We treated parent A549 with 25 μM cisplatin for 96 hours followed by at least 14 days of recovery to generate Cis-R A549 cells. Interestingly, Cis-R spheroids exhibited significantly lower rates of proliferation (P<0.01) relative to parent A549 spheroids as measured by Ki67 index. Cis-R cells and spheroids exhibit increased genetic expression for genes involved in inflammation, EMT, proliferation and chemoresistance. Furthermore, Cis-R cells in 2D displayed a mesenchymal-like morphology and further exhibited dysmorphic spheroid morphology in 3D. We then developed a membrane free microphysiological system (MPS) comprised of two adjacent and contiguous tissue layers to investigate cancer mediated angiogenesis. We seeded one layer with lung fibroblasts and endothelial cells, which formed a vascularized network, while the other lane was cultured with no cells (control), parent A549 spheroids, or Cis-R A549 spheroids. Media void of VEGF was used throughout the duration of the experiment to isolate the effect of TME-derived factors. We measured increased angiogenetic sprouting induced by Cis-R tissues relative to parent using the following metrics: sprout density (Cis-R = 49.86 ± 10.93 µm/µm2, parent = 13.73 ± 4.12 µm/µm2, blank = 14.76 ± 4.22 µm/µm2), sprout length (Cis-R = 15006 ± 5507 µm, parent = 2466 ± 639.1 µm, blank = 4749 ± 1547 µm), and sprout area fraction (Cis-R = 0.10 ± 0.028, parent = 0.041 ± 0.010, blank = 0.032 ± 0.0095). Thus, the creation of drug resistant tumor cell sub lines enables the construction of more aggressive engineered tumor microenvironments. Future iterations of our MPS model will be used to investigate angiogenesis in the context of additional cancers, including breast and colon, while transitioning to the use of patient-derived organoids. Citation Format: Elisabet Olsen, G. Wills Kpeli, Omar M.K. Ahmad, Mark Mondrinos. Increased sprouting angiogenesis in a microphysiological model of cisplatin-resistant recurrent lung cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5386.