An Immunogenic Model of KRAS-Mutant Lung Cancer Enables Evaluation of Targeted Therapy and Immunotherapy Combinations

Boumelha, Jesse; Trécesson, Sophie de Carné; Law, Emily K.; Romero-Clavijo, Pablo; Coelho, Matthew A.; Ng, Kevin W.; Mugarza, Edurne; Moore, Christopher; Rana, Sareena; Caswell, Deborah R.; Murillo, Miguel; Hancock, David C.; Argyris, Prokopios P.; Brown, William L.; Durfee, Cameron; Larson, Lindsay K.; Suárez‐Bonnet, Alejandro; Priestnall, Simon L.; East, Philip; Ross, Sarah J.; Kassiotis, George; Molina‐Arcas, Míriam; Swanton, Charles; Harris, Reuben S.; Downward, Julian

doi:10.1158/0008-5472.can-22-0325

Cited by 35 publications

(92 citation statements)

References 46 publications

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“…We did not find any macroscopic evidence of reproducible changes in any organs that would be consistent with ADAR1 overexpression, or its individual isoforms p110 or p150, leading to formation of tumors in vivo. It has been demonstrated using analogous models that the C-to-U base editor APOBEC3 will induce tumors in mice following overexpression in vivo, demonstrating that such systems will yield in vivo cancer if the overexpressed protein can function as an oncogene (94)(95)(96). Based on the two models we proposed, we did not find evidence to support the conclusion that ADAR1 was functioning as an oncogene.…”

Section: Discussioncontrasting

confidence: 56%

Overexpression of ADAR1 in mice does not initiate or accelerate cancer formationin vivo

Ruiz

Chalk

Goradia

et al. 2023

Preprint

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Adenosine to inosine editing (A-to-I) in regions of double stranded RNA (dsRNA) is mediated by adenosine deaminase acting on RNA 1 (ADAR1) or ADAR2. ADAR1 and A-to-I editing levels are increased in many human cancers. It is not established if elevated ADAR1 represents a driver or passenger during cancer formation. We established a series of murine alleles to allow in vivo overexpression of ADAR1, its individual isoforms or mutant forms of ADAR1 to understand how it contributes to cancer pathogenesis. The widespread overexpression of ADAR1 or either the p110 or p150 isoforms as sole lesions was well tolerated and did not result in cancer formation. Therefore, ADAR1 overexpression alone is not sufficient to initiate cancer. We demonstrate that endogenous ADAR1 and A-to-I editing levels increased upon immortalization by loss of p53 in murine cells, consistent with the observations from human cancers. We tested if ADAR1 overexpression could co-operate with cancer initiated by loss of tumour suppressors using a model of osteosarcoma. We did not see a disease potentiating or modifying effect of overexpressing ADAR1 or its isoforms. We conclude that the increase in ADAR1 expression and A-to-I editing in cancers is a passenger, rather than a driver, of tumor formation.

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Section: Discussioncontrasting

confidence: 56%

Overexpression of ADAR1 in mice does not initiate or accelerate cancer formationin vivo

Ruiz

Chalk

Goradia

et al. 2023

Preprint

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“…Similarly, in lung cancer, SHP2 inhibitors induce CXCR2 ligands that recruit myeloid derived suppressor cells and limit the response to this therapy 18 . Multiple groups including ours have recently demonstrated the involvement of anti-tumor immunity in the therapeutic response to KRAS G12C inhibitors 10,12,13,15 . We recently reported the involvement of adaptive immunity in the therapeutic response of murine lung cancer cells driven by oncogenic EGFR to the TKI, osimertinib 14 .…”

Section: Quantification Of Lymphocytes and Pmns In Alk+ Patient Biopsiesmentioning

confidence: 96%

“…Our recent published studies demonstrate that EGFR-targeted inhibitors induce an interferon (IFN) response program that varies markedly between distinct EGFR mutant lung cancer cell lines and positively associates with the duration of therapeutic response in EGFR-mutant lung cancer patients 7 . In fact, there is a growing literature supporting the role of host immune cells in overall therapeutic response to precision oncology agents and cytotoxic drugs [8][9][10][11][12][13][14][15][16][17][18][19] . However, the mechanisms whereby TKIs induce factors mediating paracrine signaling to the immune microenvironment, and the contribution to the overall therapeutic response are not well understood.…”

Section: Introductionmentioning

confidence: 99%

Durable responses to alectinib in murine models of EML4-ALK lung cancer requires adaptive immunity

et al. 2023

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Lung cancers bearing oncogenic EML4-ALK fusions respond to targeted tyrosine kinase inhibitors (TKIs; e.g., alectinib), with variation in the degree of shrinkage and duration of treatment (DOT). However, factors that control this response are not well understood. While the contribution of the immune system in mediating the response to immunotherapy has been extensively investigated, less is known regarding the contribution of immunity to TKI therapeutic responses. We previously demonstrated a positive association of a TKI-induced interferon gamma (IFNγ) transcriptional response with DOT in EGFR-mutant lung cancers. Herein, we used three murine models of EML4-ALK lung cancer to test the role for host immunity in the alectinib therapeutic response. The cell lines (EA1, EA2, EA3) were propagated orthotopically in the lungs of immunocompetent and immunodeficient mice and treated with alectinib. Tumor volumes were serially measured by μCT and immune cell content was measured by flow cytometry and multispectral immunofluorescence. Transcriptional responses to alectinib were assessed by RNAseq and secreted chemokines were measured by ELISA. All cell lines were similarly sensitive to alectinib in vitro and as orthotopic tumors in immunocompetent mice, exhibited durable shrinkage. However, in immunodeficient mice, all tumor models rapidly progressed on TKI therapy. In immunocompetent mice, EA2 tumors exhibited a complete response, whereas EA1 and EA3 tumors retained residual disease that rapidly progressed upon termination of TKI treatment. Prior to treatment, EA2 tumors had greater numbers of CD8+ T cells and fewer neutrophils compared to EA1 tumors. Also, RNAseq of cancer cells recovered from untreated tumors revealed elevated levels of CXCL9 and 10 in EA2 tumors, and higher levels of CXCL1 and 2 in EA1 tumors. Analysis of pre-treatment patient biopsies from ALK+ tumors revealed an association of neutrophil content with shorter time to progression. Combined, these data support a role for adaptive immunity in durability of TKI responses and demonstrate that the immune cell composition of the tumor microenvironment is predictive of response to alectinib therapy.

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“…Previous studies have demonstrated that KRAS-SHP2-MAP kinase pathway-targeted agents induce expression of multiple chemokines in KRAS-mutant lung cancer cell lines ( 21 , 44 , 45 ) and may provide a mechanism by which oncogene-targeted drugs induce cross-talk between cancer cells and the tumor microenvironment. Interrogation of RNAseq data generated from LLC 46 NRAS KO, CMT-KRAS-G12C.54 and mKRC.1 cells treated in vitro for 6 hr to 3 days with 30 nM MRTX-1257 reveals distinct expression patterns of chemokines with defined anti- and pro-tumorigenic functions ( 46 ) ( Figure 6A ).…”

Section: Resultsmentioning

confidence: 99%

“…The murine KRAS G12C -driven lung cancer cell lines developed in this study provide novel models to investigate KRAS G12C inhibitor responsiveness in the immune competent setting. This is critical since oncogene-targeted agents including KRAS G12C inhibitors have been shown to initiate functional interactions with host immunity ( 27 , 28 , 34 , 44 , 45 ). The new lung cancer cell lines described herein uniformly exhibit high in vitro sensitivity to KRAS G12C inhibitors and altered growth signaling in CMT KRAS G12C clones compared to parental G12V cells.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of KRASG12C inhibitor responses in novel murine KRASG12C lung cancer cell line models

Sisler

Hinz²,

Le³

et al. 2023

Front. Oncol.

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IntroductionThe KRAS(G12C) mutation is the most common genetic mutation in North American lung adenocarcinoma patients. Recently, direct inhibitors of the KRASG12C protein have been developed and demonstrate clinical response rates of 37-43%. Importantly, these agents fail to generate durable therapeutic responses with median progression-free survival of ~6.5 months.MethodsTo provide models for further preclinical improvement of these inhibitors, we generated three novel murine KRASG12C-driven lung cancer cell lines. The co-occurring NRASQ61L mutation in KRASG12C-positive LLC cells was deleted and the KRASG12V allele in CMT167 cells was edited to KRASG12C with CRISPR/Cas9 methods. Also, a novel murine KRASG12C line, mKRC.1, was established from a tumor generated in a genetically-engineered mouse model.ResultsThe three lines exhibit similar in vitro sensitivities to KRASG12C inhibitors (MRTX-1257, MRTX-849, AMG-510), but distinct in vivo responses to MRTX-849 ranging from progressive growth with orthotopic LLC-NRAS KO tumors to modest shrinkage with mKRC.1 tumors. All three cell lines exhibited synergistic in vitro growth inhibition with combinations of MRTX-1257 and the SHP2/PTPN11 inhibitor, RMC-4550. Moreover, treatment with a MRTX-849/RMC-4550 combination yielded transient tumor shrinkage in orthotopic LLC-NRAS KO tumors propagated in syngeneic mice and durable shrinkage of mKRC.1 tumors. Notably, single-agent MRTX-849 activity in mKRC.1 tumors and the combination response in LLC-NRAS KO tumors was lost when the experiments were performed in athymic nu/nu mice, supporting a growing literature demonstrating a role for adaptive immunity in the response to this class of drugs.DiscussionThese new models of murine KRASG12C mutant lung cancer should prove valuable for identifying improved therapeutic combination strategies with KRASG12C inhibitors.

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An Immunogenic Model of KRAS-Mutant Lung Cancer Enables Evaluation of Targeted Therapy and Immunotherapy Combinations

Cited by 35 publications

References 46 publications

Overexpression of ADAR1 in mice does not initiate or accelerate cancer formationin vivo

Overexpression of ADAR1 in mice does not initiate or accelerate cancer formationin vivo

Durable responses to alectinib in murine models of EML4-ALK lung cancer requires adaptive immunity

Evaluation of KRASG12C inhibitor responses in novel murine KRASG12C lung cancer cell line models

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