PO.ET03.05 · 实验与分子治疗
Multi-lineage evolution of drug resistance via a novel keratin 17+ drug tolerant persister population in EGFR -mutant NSCLC
作者与单位
摘要 Abstract
Background: EGFR mutant lung cancers initially respond to frontline targeted therapy. However, resistance inevitably develops through diverse mechanisms. Resistant tumors universally emerge from drug tolerant persister cells (DTPCs) that survive initial treatment. Despite this knowledge, the biology of EGFR mutant DTPCs is not well understood. This study was conducted to interrogate DTPC phenotypes that enable evolution of therapy resistance.
Methods: DTPC models were generated by treating EGFR mutant cell lines with >IC90 concentrations of osimertinib for 14 days. After 14 days, cells were profiled with 10X 3' single cell RNA sequencing (scRNAseq). Seurat was used to integrate data, normalize data, cluster cells, and conduct differential expression analyses. UCell was used to score cells for activity of biology pathways. scanpy was used to analyze scRNAseq data from the Gardner et al. ERPMT mouse model. Human patient samples were collected from lung cancer patients treated at MD Anderson Cancer Center using IRB approved protocols. Samples were profiled with 10X 5' scRNAseq. scanpy was used to integrate data, normalize data, and cluster cells. Palantir was used to perform trajectory analyses.
Results: Differential expression (DE) analysis comparing DTPCs and control cells were conducted for cell line pairs. Using DE results, we generated consensus gene signatures capturing genes universally upregulated or downregulated across DTPC models. Review of these genes identified that KRT17 was highly upregulated in osimertinib DTPCs. We found KRT17 + DTPCs upregulate epithelial-to-mesenchymal (EMT), stemness, and aberrant basaloid signatures compared to KRT17 - DTPCs. Subclustering showed that KRT17 DTPCs are heterogeneous and label non-overlapping EMT, proliferative, and MET expressing populations. To validate our in vitro findings, we analyzed scRNAseq datasets from multiple in vivo sources. In GEMM models, Krt17 + cells populate minimal residual disease (MRD) following suppression of mutant EGFR. Krt17 + cells were a stem-like population that emerged as residual tumor cells lost alveolar epithelial identity prior to neuroendocrine transformation. Lastly, we confirmed the clinical relevance of our findings by investigating human patient clinical specimens. Compared to treatment naïve samples, KRT17 + cells were significantly enriched in osimertinib-treated MRD samples. Trajectory analyses demonstrated that KRT17 expression characterized cell populations that preceded development of resistance through several known mechanisms, including MET amplification and histologic transformation (HT).
Conclusions: Our data demonstrate that EGFR-mutant cells surviving initial treatment converge onto a novel KRT17 + DTPC state which serves as a multipotent progenitor population capable of leveraging diverse mechanisms to resist therapy, including HT.
利益披露 Disclosure
B. B. Morris, None.
M. B. Nilsson,
Spectrum Pharmaceuticals Other, MN receives licensing fees and royalties from spectrum pharmaceuticals.
E. Earlie, None..
E. E. Gardner, None..
H. Chen, None..
S. G. Trevino, None..
A. J. Halliday, None..
Y. Xi, None..
J. Wang, None.
N. Vokes,
AstraZeneca ), Other, Consultant/Advisory Board.
Guardant Other, Consulting/Advisory Board; Honoraria.
Bristol Myers Squibb ), Other, Trial support.
Catalyst Pharmaceuticals Other, Consulting/Advisory Board.
ImmunityBio Other, Consulting/Advisory Board.
Sanofi ), Other, Trial support.
Mirati ).
Circulogene ), Other, Funding.
Pfizer Other, Consulting/Advisory Board.
Summit Therapeutics Other, Consulting/Advisory Board; Trial support.
OncoHost Other, Consulting/Advisory Board; Trial support.
Regeneron Other, Trial support; travel reimbursement.
Amgen Other, Trial support.
IDEAYA Other, Trial support.
EMD serono Other, Trial support.
Dava Onc Other, Honoraria.
Cardinal Health Other, Honoraria.
MJH Other, Honoraria.
Medlive Other, Honoraria.
HMP Global Other, Honoraria.
D. Gibbons,
Menarini Ricerche Other, Scientific Advisory Board.
Onconova Other, Scientific Advisory Board.
Aktis Oncology Other, Scientific Advisory Board.
Eli Lilly ), Other, Scientific Advisory Board.
Ideology Health Other, Honoraria.
NGM Biopharmaceuticals ).
Boehringer Ingelheim ).
Mirati ).
Bristol Myers Squibb ).
J. Zhang,
Merck ).
Johnson and Johnson ), Other, Consulting.
Norvartis ), Other, Consulting.
Summit Therapeutics ).
Helius ).
AstraZeneca Other, Consulting.
GenePlus Other, Consulting.
OrigMed Other, Consulting.
Innovent Other, Consulting.
Varian Other, Consulting.
Catalyst Therapeutics Other, Consulting.
A. M. Laughney, None.
Y. Y. Elamin,
AstraZeneca ), Other, Advisory Board.
Eli Lilly Other, Advisory Board.
Norvartis Other, Advisory Board.
Sanofi Other, Advisory Board.
Takeda ), Other, Advisory Board.
Bristol Myers Squibb Other, Advisory Board.
Nuvation ), Other, Advisory Board.
Mirati Other, Advisory Board.
Merus ), Other, Advisory Board.
Tahioo ), Other, Advisory Board.
BluePrint Medicines Other, Advisory Board.
Catalyst Pharmaceuticals Other, Advisory Board.
Johnson & Johnson Other, Advisory Board.
Kestrel Therapeutics Other, Advisory Board.
Telligene ).
Ellipses ).
X. Le,
ArriVent ), Other, Consulting.
Bayer AG ), Other, Consulting.
Boehringer Ingelheim ), Other, Consulting.
Eli Lilly ), Other, Consulting.
EMD Serono ), Other, Consulting.
Janssen ), Other, Consulting.
Dizal ), Other, Consulting.
Regeneron ), Other, Consulting.
Takeda ).
Teligene ), Other, Consulting.
ThermoFisher ).
AstraZeneca Other, Consulting.
AbbVie Other, Consulting.
Abion Other, Consulting.
Allist Other, Consulting.
Bristol Myers Squibb Other, Consulting.
Daiichi-Sankyo Other, Consulting.
BioNTech Other, Consulting.
Roche/Genentech Other, Consulting.
Taiho Other, Consulting.
J. V. Heymach,
AstraZeneca ), Other, Advisory Board.
Boehringer Ingelheim ), Other, Advisory Board.
Bristol Myers Squibb ), Other, Advisory Board.
Mirati ), Other, Advisory Board.
Takeda ), Other, Advisory Board.
Taiho ).
Tenaci-T Therapeutics Other, Business ownership.
Spectrum Pharmaceuticals Other, Licensing/royalties.
Clinical Care Targeted Communications Other, Speaking event.
Physicians Education Resource Other, Speaking event.
Prime Education Other, Speaking event.
Bayer Other, Advisory Board.
Eli Lilly Other, Advisory Board.
Janssen Pharmaceuticals Other, Advisory Board.
Jazz Pharmaceuticals Other, Advisory Board.
OncoHost Other, Advisory Board.
BioNTech AG Other, Advisory Board.
Curio Science Other, Advisory Board.
DAVA Oncology Other, Advisory Board.
EMD Serono Other, Advisory Board.