TY - JOUR
T1 - Tumorigenic circulating tumor cells from xenograft mouse models of non-metastatic NSCLC patients reveal distinct single cell heterogeneity and drug responses
AU - Suvilesh, Kanve N.
AU - Nussbaum, Yulia I.
AU - Radhakrishnan, Vijay
AU - Manjunath, Yariswamy
AU - Avella, Diego M.
AU - Staveley-O’Carroll, Kevin F.
AU - Kimchi, Eric T.
AU - Chaudhuri, Aadel A.
AU - Shyu, Chi Ren
AU - Li, Guangfu
AU - Pantel, Klaus
AU - Warren, Wesley C.
AU - Mitchem, Jonathan B.
AU - Kaifi, Jussuf T.
N1 - Funding Information:
This study was supported by a Mizzou Advantage Interdisciplinary Research Grant (MAIRG) (J.T.K., G.L.). J.B.M. received funding from the Department of Veterans Affairs K2BX004346-01A1. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Department of Veterans Affairs. The funding bodies had no role in study design, collection, analysis, interpretation of data or writing the manuscript.
Funding Information:
We are exceedingly grateful to all patients for their voluntary participation. The authors thank Christopher Bottoms (MU Informatics Research Core) for helping with data analysis, Nathan Bivens (MU DNA Core) for suggestions and guidance on sample preparation for snRNA-seq, and David Pittman (MU Department of Pathology) for reviewing histopathological and immunohistochemical staining. We thank Nancy Walker (MU Department of Surgery) for critical review of the manuscript. All raw single-nuclear RNA sequencing data were deposited in NCBI Sequence Read Archive (SRA) and are accessible through project number PRJNA755249. To validate the snRNA-seq results, we utilized open access scRNA-seq data that were downloaded using the accession number GSE123904. Codes used for data analysis are deposited in Code Ocean and available for download at (https://codeocean.com/capsule/7961492/tree). All other datasets generated and analyzed in the current study are available from the corresponding authors upon request.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Background: Circulating tumor cells (CTCs) are liquid biopsies that represent micrometastatic disease and may offer unique insights into future recurrences in non-small cell lung cancer (NSCLC). Due to CTC rarity and limited stability, no stable CTC-derived xenograft (CDX) models have ever been generated from non-metastatic NSCLC patients directly. Alternative strategies are needed to molecularly characterize CTCs and means of potential future metastases in this potentially curable patient group. Methods: Surgically resected NSCLC primary tumor tissues from non-metastatic patients were implanted subcutaneously in immunodeficient mice to establish primary tumor patient-derived xenograft (ptPDX) models. CTCs were isolated as liquid biopsies from the blood of ptPDX mice and re-implanted subcutaneously into naïve immunodeficient mice to generate liquid biopsy CTC-derived xenograft (CDX) tumor models. Single cell RNA sequencing was performed and validated in an external dataset of non-xenografted human NSCLC primary tumor and metastases tissues. Drug response testing in CDX models was performed with standard of care chemotherapy (carboplatin/paclitaxel). Blockade of MYC, which has a known role in drug resistance, was performed with a MYC/MAX dimerization inhibitor (10058-F4). Results: Out of ten ptPDX, two (20%) stable liquid biopsy CDX mouse models were generated. Single cell RNA sequencing analysis revealed an additional regenerative alveolar epithelial type II (AT2)-like cell population in CDX tumors that was also identified in non-xenografted NSCLC patients’ metastases tissues. Drug testing using these CDX models revealed different treatment responses to carboplatin/paclitaxel. MYC target genes and c-MYC protein were upregulated in the chemoresistant CDX model, while MYC/MAX dimerization blocking could overcome chemoresistance to carboplatin/paclitaxel. Conclusions: To overcome the lack of liquid biopsy CDX models from non-metastatic NSCLC patients, CDX models can be generated with CTCs from ptPDX models that were originally established from patients’ primary tumors. Single cell analyses can identify distinct drug responses and cell heterogeneities in CDX tumors that can be validated in NSCLC metastases tissues. CDX models deserve further development and study to discover personalized strategies against micrometastases in non-metastatic NSCLC patients.
AB - Background: Circulating tumor cells (CTCs) are liquid biopsies that represent micrometastatic disease and may offer unique insights into future recurrences in non-small cell lung cancer (NSCLC). Due to CTC rarity and limited stability, no stable CTC-derived xenograft (CDX) models have ever been generated from non-metastatic NSCLC patients directly. Alternative strategies are needed to molecularly characterize CTCs and means of potential future metastases in this potentially curable patient group. Methods: Surgically resected NSCLC primary tumor tissues from non-metastatic patients were implanted subcutaneously in immunodeficient mice to establish primary tumor patient-derived xenograft (ptPDX) models. CTCs were isolated as liquid biopsies from the blood of ptPDX mice and re-implanted subcutaneously into naïve immunodeficient mice to generate liquid biopsy CTC-derived xenograft (CDX) tumor models. Single cell RNA sequencing was performed and validated in an external dataset of non-xenografted human NSCLC primary tumor and metastases tissues. Drug response testing in CDX models was performed with standard of care chemotherapy (carboplatin/paclitaxel). Blockade of MYC, which has a known role in drug resistance, was performed with a MYC/MAX dimerization inhibitor (10058-F4). Results: Out of ten ptPDX, two (20%) stable liquid biopsy CDX mouse models were generated. Single cell RNA sequencing analysis revealed an additional regenerative alveolar epithelial type II (AT2)-like cell population in CDX tumors that was also identified in non-xenografted NSCLC patients’ metastases tissues. Drug testing using these CDX models revealed different treatment responses to carboplatin/paclitaxel. MYC target genes and c-MYC protein were upregulated in the chemoresistant CDX model, while MYC/MAX dimerization blocking could overcome chemoresistance to carboplatin/paclitaxel. Conclusions: To overcome the lack of liquid biopsy CDX models from non-metastatic NSCLC patients, CDX models can be generated with CTCs from ptPDX models that were originally established from patients’ primary tumors. Single cell analyses can identify distinct drug responses and cell heterogeneities in CDX tumors that can be validated in NSCLC metastases tissues. CDX models deserve further development and study to discover personalized strategies against micrometastases in non-metastatic NSCLC patients.
UR - http://www.scopus.com/inward/record.url?scp=85126262480&partnerID=8YFLogxK
U2 - 10.1186/s12943-022-01553-5
DO - 10.1186/s12943-022-01553-5
M3 - Letter
C2 - 35279152
AN - SCOPUS:85126262480
SN - 1476-4598
VL - 21
JO - Molecular Cancer
JF - Molecular Cancer
IS - 1
M1 - 73
ER -