|
| Status |
Public on Jan 24, 2024 |
| Title |
Extracellular niche and tumor microenvironment enhance the impact of KRAS inhibitors in pancreatic cancer models [CRISPR] |
| Organism |
Homo sapiens |
| Experiment type |
Other
|
| Summary |
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease for which new therapeutic interventions are needed. Here, we assessed the cellular response to pharmacological KRAS inhibition, targeting the central oncogenic factor in PDAC. In a panel of PDAC cell lines, pharmaceutical inhibition of the KRASG12D allele with MRTX1133 yields variable efficacy in the suppression of cell growth and downstream gene expression programs in 2D culture. CRISPR-Cas9 loss-of-function screens identified ITGB1 as a novel driver for enhanced therapeutic response that regulates mechanotransdcution signaling and YAP/TAZ expression, which is further confirmed by gene specific knockdowns and combinatorial drug synergy. Interestingly, MRTX1133 is considerably more efficacious in the context of 3D cell cultures. Moreover, MRTX1133 elicits a more pronounced cytostatic effect in controlling the in vivo tumor growth in PDAC patient-derived xenografts. In syngeneic models, KRASG12D inhibition leads to potent tumor regression that did not occur in immune-deficient hosts. Digital spatial profiling on tumor tissues indicates that MRTX1133-mediated KRAS inhibition enhances interferon-γ signaling and induces antigen presentation that modulates the tumor microenvironment. Further investigation on the immunological response using single-cell sequencing and multispectral imaging reveals that tumor regression is associated with suppression of neutrophils and influx of effector CD8+ T-cells. Thus, both tumor cell-intrinsic and extrinsic events contribute to response and credential KRASG12D inhibition as a promising strategy for a large percentage of PDAC tumors.
|
| |
|
| Overall design |
ASPC1 and UM53 cells were infected with the TKOv3 lentiviral particles at an MOI ~0.3 and the positive clones were selected using puromycin to achieve a mutant pool comprising at least 200-fold coverage of the library. The resulting mutant pool was grown in the absence and presence of KRAS inhibitors, which include MRTX1133 (6.25 nM) and MRTX849 ( 500 nM) for at least 5 passages. Genomic DNA was extracted and subjected to sequencing library construction by amplifying the gRNA inserts
|
| |
|
| Contributor(s) |
Kumarasamy V, Wang J, Frangou C, Wan Y, Dynka A, Rosenheck H, Dey P, Abel EV, Knudsen ES, Witkiewicz AK |
| Citation(s) |
38294344 |
| |
| Submission date |
Dec 06, 2023 |
| Last update date |
Apr 25, 2024 |
| Contact name |
Jianxin Wang |
| Organization name |
Roswell Park Comprehensive Cancer Center
|
| Department |
Molecular and Cellular Biology
|
| Street address |
665 Elm Street
|
| City |
Buffalo |
| State/province |
NY |
| ZIP/Postal code |
14203 |
| Country |
USA |
| |
|
| Platforms (1) |
|
| Samples (10)
|
|
| This SubSeries is part of SuperSeries: |
| GSE249541 |
Extracellular niche and tumor microenvironment enhance the impact of KRAS inhibitors in pancreatic cancer models. |
|
| Relations |
| BioProject |
PRJNA1049457 |
Less...
More...