AUTHOR=Andus Ingo , Prall Friedrich , Linnebacher Michael , Linnebacher Christina S. 

TITLE=Establishment, characterization, and drug screening of low-passage patient individual non-small cell lung cancer in vitro models including the rare pleomorphic subentity

JOURNAL=Frontiers in Oncology

VOLUME=Volume 13 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2023.1089681

DOI=10.3389/fonc.2023.1089681

ISSN=2234-943X

ABSTRACT=Introduction
For pre-clinical drug development and precision oncology research, robust cancer cell models are essential. Patient-derived models in low passages retain more genetic and phenotypic characteristics of their original tumors than conventional cancer cell lines. Subentity, individual genetics and heterogeneity influence drug sensitivity and clinical outcome greatly.
Material and Methods
Here we report on establishment and characterization of three patient derived cell lines (PDC) of different subentities of non-small cell lung cancer (NSCLC): adeno-, squamous cell and pleomorphic carcinoma. The in-depth characterization of our PDC included phenotype, proliferation, surface protein expression, invasion and migration behavior as well as whole-exome and RNA sequencing. Additionally, in vitro drug sensitivity towards standard of care chemotherapeutic regimens were evaluated.
Results
The pathological and molecular properties of the patients´ tumors were preserved in the PDC models HROLu22, HROLu55 and HROBML01. All cell lines expressed HLA I, while none were positive for HLA II. The epithelial cell marker CD326 and the lung tumor markers CCDC59, LYPD3 and DSG3 were also detected. The most frequently mutated genes included TP53, MXRA5, MUC16 and MUC19. Among the most over expressed genes in tumor cells compared to normal tissue were the transcription factors HOXB9, SIM2, ZIC5, SP8, TFAP2A, FOXE1, HOXB13 and SALL4, the cancer testis antigen CT83 and the cytokine IL23A. The most down-regulated genes on the RNA level encode the long non-coding RNA LANCL1-AS1, LINC00670, BANCR, and LOC100652999, the regulator of angiogenesis ANGPT4, the signaling molecules PLA2G1B and RS1 as well as the immune modulator SFTPD. Furthermore, neither pre-existing therapy resistances nor drug antagonistic effects could be observed.
Conclusion
In summary, we successfully established three novel NSCLC PDC models from an adeno-, a squamous cell and a pleomorphic carcinoma. Of note, this is the first description of a NSCLC cell model of the pleomorphic subentity. The detailed characterization including molecular, morphological and drug-sensitivity profiling makes these models valuable pre-clinical tools for drug development applications and research on precision cancer therapy. The pleomorphic model additionally enables research on a functional and cell based level of this rare NCSLC subentity for the first time.