Population-wide gene disruption in the murine lung epithelium via AAV-mediated delivery of CRISPR-Cas9 components

Chen, Honglin; Durinck, Steffen; Patel, Hetal; Foreman, Oded; Mesh, Kathryn; Eastham, Jeffrey; Caothien, Roger; Newman, Robert J.; Roose‐Girma, Merone; Darmanis, Spyridon; Warming, Søren; Lattanzi, Annalisa; Liang, Yuxin; Haley, Benjamin

doi:10.1016/j.omtm.2022.10.016

Cited by 8 publications

(3 citation statements)

References 83 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Animal rooms were temperature and humidity-controlled, between 68 to 79°F (20.0 to 26.1°C) and 30 to 70% respectively, with 10 to 15 room air exchanges per hour. Rosa26.Cas9.HTN.ki.B6N.4B3-1 (Cas9) mice were generated and maintained at Genentech (28). Heterozygous and homozygous knock-in Cas9 mice of both sexes were used.…”

Section: Animal Studiesmentioning

confidence: 99%

RIT1M90I is a driver of lung adenocarcinoma tumorigenesis and resistance to targeted therapy

Maddalo,

DiMarco,

Ravicahndran

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

RIT1 is a RAS-family GTPase that is mutated in 2.4% and amplified in up to 14% of lung adenocarcinoma patients. Yet, the oncogenic potential of RIT1 in the lungs is not completely understood. Many patients with RIT1 alterations are considered ‘oncogene-negative’, hence they are not eligible for any targeted therapy in the clinic. The role of RIT1 in cancer has been historically understudied due to the lack of in vitro and in vivo models harboring RIT1 alterations. In this study, we generated the first murine model of RIT1M90I-mutant lung cancer. We found that RIT1M90I was tumorigenic in the lungs and the tumors displayed histopathological features similar to lung adenocarcinoma. We leveraged this new model to assess both ex vivo and in vivo sensitivity of RIT1 tumors to the SHP2 inhibitor, migoprotafib, in combination with other targeted therapies. Finally, we showed that RIT1M90I can also drive resistance to the KRASG12C inhibitor, divarasib, and that the combination with migoprotafib can revert this phenotype. Our data shows that RIT1M90I is a bona fide oncogenic driver of NSCLC and mediator of targeted therapy resistance as a co-occurring mutation. Taken together, our study suggests that RIT1-altered cancer patients may benefit from combination treatment with SHP2 inhibitor in the primary and resistance setting.

show abstract

Section: Animal Studiesmentioning

confidence: 99%

RIT1M90I is a driver of lung adenocarcinoma tumorigenesis and resistance to targeted therapy

Maddalo,

DiMarco,

Ravicahndran

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…AAV2 (Upstaza, to treat AADC deficiency; AADC gene) 12 Heart AAV9, [178][179][180][181][182][183][184][185] AAVMYO 186 -Kidney AAV2, 187 AAV6, 188 AAV8, 189,190 AAV6.2 190 -Cochlear cells AAV9, 191 AAV-PHP.B, [192][193][194][195][196] AAV-Anc80L65, 191,[197][198][199]200,201 -Lungs AAV1, [202][203][204] AAV6, [205][206][207] AAV6.2FF, 208,209 Abbreviation: AAV, adeno-associated viral vector.…”

Section: Immune Response To Aavmentioning

confidence: 99%

The menace of severe adverse events and deaths associated with viral gene therapy and its potential solution

Kachanov,

Kostyusheva,

Brezgin

et al. 2024

Medicinal Research Reviews

View full text Add to dashboard Cite

Over the past decade, in vivo gene replacement therapy has significantly advanced, resulting in market approval of numerous therapeutics predominantly relying on adeno‐associated viral vectors (AAV). While viral vectors have undeniably addressed several critical healthcare challenges, their clinical application has unveiled a range of limitations and safety concerns. This review highlights the emerging challenges in the field of gene therapy. At first, we discuss both the role of biological barriers in viral gene therapy with a focus on AAVs, and review current landscape of in vivo human gene therapy. We delineate advantages and disadvantages of AAVs as gene delivery vehicles, mostly from the safety perspective (hepatotoxicity, cardiotoxicity, neurotoxicity, inflammatory responses etc.), and outline the mechanisms of adverse events in response to AAV. Contribution of every aspect of AAV vectors (genomic structure, capsid proteins) and host responses to injected AAV is considered and substantiated by basic, translational and clinical studies. The updated evaluation of recent AAV clinical trials and current medical experience clearly shows the risks of AAVs that sometimes overshadow the hopes for curing a hereditary disease. At last, a set of established and new molecular and nanotechnology tools and approaches are provided as potential solutions for mitigating or eliminating side effects. The increasing number of severe adverse reactions and, sadly deaths, demands decisive actions to resolve the issue of immune responses and extremely high doses of viral vectors used for gene therapy. In response to these challenges, various strategies are under development, including approaches aimed at augmenting characteristics of viral vectors and others focused on creating secure and efficacious non‐viral vectors. This comprehensive review offers an overarching perspective on the present state of gene therapy utilizing both viral and non‐viral vectors.

show abstract

“…Immortalized macrophages (ER-Hoxb8-immortalized murine myeloid progenitor cells 61,62 ) were established from Cas9 transgenic mice 63 . Cells were cultured in RPMI 1640 medium supplemented with 10% (v/v) FBS, 20 ng/mL murine granulocyte-macrophage colony-stimulating factor (GM-CSF, eBioscience), and 1 mM b-estradiol (Millipore Sigma).…”

Section: Cell Culturementioning

confidence: 99%

Highly multiplexed, image-based pooled screens in primary cells and tissues with PerturbView

Kudo,

Meireles,

Moncada

et al. 2023

Preprint

View full text Add to dashboard Cite

Optical pooled screening (OPS) is a highly scalable method for linking image-based phenotypes with cellular perturbations. However, it has thus far been restricted to relatively low-plex phenotypic readouts in cancer cell lines in culture, due to limitations associated within situsequencing (ISS) of perturbation barcodes. Here, we developed PerturbView, an OPS technology that leveragesin vitrotranscription (IVT) to amplify barcodes prior to ISS, enabling screens with highly multiplexed phenotypic readouts across diverse systems, including primary cells and tissues. We demonstrate PerturbView in iPSC-derived neurons, primary immune cells, and tumor tissue sections from animal models. In a screen of immune signaling pathways in primary bone marrow-derived macrophages, PerturbView uncovered both known and novel regulators of NFκB signaling. Furthermore, we combined PerturbView with spatial transcriptomics in tissue sections from a mouse xenograft model, paving the way toin vivoscreens with rich optical and transcriptomic phenotypes. PerturbView broadens the scope of OPS to a wide range of models and applications.

show abstract

Population-wide gene disruption in the murine lung epithelium via AAV-mediated delivery of CRISPR-Cas9 components

Cited by 8 publications

References 83 publications

RIT1M90I is a driver of lung adenocarcinoma tumorigenesis and resistance to targeted therapy

RIT1M90I is a driver of lung adenocarcinoma tumorigenesis and resistance to targeted therapy

The menace of severe adverse events and deaths associated with viral gene therapy and its potential solution

Highly multiplexed, image-based pooled screens in primary cells and tissues with PerturbView

Contact Info

Product

Resources

About