A mouse model for the study of anti-tumor T cell responses in Kras-driven lung adenocarcinoma

Fitzgerald, Brittany; Connolly, Kelli A.; Cui, Can; Fagerberg, Eric; Mariuzza, Dylan L.; Hornick, Noah I.; Foster, Gena G.; William, Ivana; Cheung, Julie F.; Joshi, Nikhil S.

doi:10.1016/j.crmeth.2021.100080

Cited by 18 publications

(12 citation statements)

References 89 publications

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“…4A). This is in agreement with previous publications, which indicated that the lack of neoantigens in the KP model results not elicit tumor-speci c T cell responses (Fitzgerald et al 2021; DuPage, Dooley, and Jacks 2009). However, a subset of tumor nodules exhibited signi cant in ltration by CD8-positive cells, a critical subpopulation of MHC class I-restricted T cells that are mediators of adaptive immunity (Figs.…”

Section: Single Tumor Nodules Induced By Low-titer Aav-cre Transducti...supporting

confidence: 93%

An optimized protocol for generating and monitoring conditional orthotopic lung cancer in the KP mouse model at BSL-1

Deng

Dubey

et al. 2023

Preprint

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Background: The inducible Kras/p53 lung adenocarcinoma mouse model, which faithfully recapitulates human disease, is routinely initiated by the intratracheal instillation of a virus-based Cre recombinase delivery system. Handling virus-based delivery systems requires elevated biosafety levels, e.g., biosafety level 2 (BSL-2). However, in experimental animal research facilities, following exposure to viral vectors in a BSL-2 environment, rodents may not be reclassified to BSL-1 according to standard practice, preventing access to small animal micro-computed tomography (micro-CT) scanners that are typically housed in general access areas such as BSL-1 rooms. Therefore, our goal was to adapt the protocol so that the Cre-induced KP mouse model could be handled under BSL-1 conditions during the entire procedure. Results: The Kras-Lox-STOP-Lox-G12D/p53 flox/flox (KP)-based lung adenocarcinoma mouse model was activated by intratracheal instillation of either a virus-based Cre delivery system or a gutless, adeno-associated, Cre-expressing vector. Tumor growth was monitored over time by micro-CT. We have successfully substituted the virus-based Cre delivery system with a commercially available, gutless, adeno-associated, Cre-expressing vector that allows the KP mouse model to be handled and imaged in a BSL-1 facility. By optimizing the anesthesia protocol and switching to a microscope-guided vector instillation procedure, productivity was increased and procedure-related complications were significantly reduced. In addition, repeated micro-CT analysis of individual animals allowed us to monitor tumor growth longitudinally, dramatically reducing the number of animals required per experiment. Conclusion: Modifications to the anesthesia and instillation protocols increased the productivity of the original KP protocol. In addition, the switch to a gutless, adeno-associated, Cre-expressing vector allowed longitudinal monitoring of tumor growth under BSL-1 conditions, significantly reducing the number of animals required for an experiment, in line with the 3R principles.

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Section: Single Tumor Nodules Induced By Low-titer Aav-cre Transducti...supporting

confidence: 93%

An optimized protocol for generating and monitoring conditional orthotopic lung cancer in the KP mouse model at BSL-1

Deng

Dubey

et al. 2023

Preprint

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“…Conversely, we found increased expression of MHC-I and MHC-II on HKP1 cells in F4.CAR-T treated mice, and observed that these remaining cells now presented reduced expression of tdTomato. We and others reported previously this can occur as a consequence of antigen editing, as cancer cells that lose or downregulate antigen are no longer under selective pressure to downregulate the antigen presentation machinery (27,39,40). Our studies therefore suggest that treatment with macrophage-targeted CAR-T promoted T-cell immunity against antigen-expressing HKP1 cell clones and selected for the outgrowth of tumor cells expressing lower amounts of this protein.…”

Section: Discussionsupporting

confidence: 61%

Targeting Macrophages with CAR T Cells Delays Solid Tumor Progression and Enhances Antitumor Immunity

Sánchez-Paulete

Mateus-Tique

Mollaoglu

et al. 2022

Cancer Immunology Research

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Tumor-associated macrophages (TAMs) are one of the most abundant cell types in many solid tumors and typically exert protumor effects. This has led to an interest in macrophage-depleting agents for cancer therapy, but approaches developed to date have had limited success in clinical trials. Here, we report the development of a strategy for TAM depletion in mouse solid tumor models using chimeric antigen receptor (CAR) T cells targeting the macrophage marker F4/80 (F4.CAR-T). F4.CAR-T cells effectively killed macrophages in vitro and in vivo without toxicity. When injected into mice bearing orthotopic lung tumors, F4.CAR-T cells infiltrated tumor lesions and delayed tumor growth comparably to PD-1 blockade, and significantly extended mouse survival. Anti-tumor effects were mediated by F4.CAR-T-produced IFN-γ, which promoted upregulation of MHC molecules on cancer cells and tumor-infiltrating myeloid cells. Notably, F4.CAR-T promoted expansion of endogenous CD8 T cells specific for tumor-associated antigen and led to immune editing of highly antigenic tumor cell clones. Antitumor impact was also observed in mouse models of ovarian and pancreatic cancer. These studies provide proof-of-principle to support CAR T cell targeting of TAMs as a means to enhance antitumor immunity.

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“…Additionally, the complex TME in GEMMs, a pertinent reflection of the patient tumor, makes it difficult to say with certainty that the impact of glutaminase inhibition in the CD8 T cells alone was responsible for their impeded function, and not a result of the cross talk within cells of the TME. Finally, given that KEAP1 and STK11 mutant lung adenocarcinoma are generally associated with cigarette smoke and high tumor mutational burden (TMB) (Ricciuti et al, 2022;Skoulidis et al, 2015), it will be important to validate these findings in murine models engineered to exhibit a higher TMB and/or neoantigens (Fitzgerald et al, 2021;Westcott et al, 2021). Despite this, our findings suggest that glutaminase function must be unimpeded in CD8 T cells to ensure that they have the ability to reach their full cytotoxic potential.…”

Section: Limitations Of the Studymentioning

confidence: 90%

Glutaminase inhibition impairs CD8 T cell activation in STK11-/Lkb1-deficient lung cancer

et al. 2022

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A mouse model for the study of anti-tumor T cell responses in Kras-driven lung adenocarcinoma

Cited by 18 publications

References 89 publications

An optimized protocol for generating and monitoring conditional orthotopic lung cancer in the KP mouse model at BSL-1

An optimized protocol for generating and monitoring conditional orthotopic lung cancer in the KP mouse model at BSL-1

Targeting Macrophages with CAR T Cells Delays Solid Tumor Progression and Enhances Antitumor Immunity

Glutaminase inhibition impairs CD8 T cell activation in STK11-/Lkb1-deficient lung cancer

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