Dataset Information

In vivo genome-wide CRISPR screens in human T cells to enhance T cell therapy for solid tumors

ABSTRACT: Large-scale CRISPR screening in human T cells holds significant promise for identifying genetic modifications that can enhance cellular immunotherapy. However, many genetic regulators of T cell performance in solid tumors may not be readily revealed in vitro. In vivo screening in tumor-bearing mice offers greater physiological relevance but has historically been limited by low intratumoral T cell recovery. Here, we developed a new model system that achieves substantially higher human T cell recovery from tumors, enabling genome-wide in vivo screens with relatively small numbers of mice. Tumor-infiltrating T cells in this model exhibit hallmarks of dysfunction compared to matched splenic T cells, creating an ideal context for screening for genetic modifiers of T cell activity in the tumor microenvironment. Using this platform, we performed two genome-wide CRISPR knockout screens to identify genes regulating T cell intratumoral abundance and effector function, as measured by IFN-γ production. The intratumoral abundance screen uncovered the P2RY8-Gα13 GPCR signaling axis as a negative regulator of human T cell infiltration into tumors. The effector function screen identified GNAS as a key regulator of T cell dysfunction in tumors. Its protein product, Gαs, acts as a convergent signaling node downstream of multiple GPCRs that sense different suppressive ligands. Targeted GNAS knockout rendered T cells resistant to multiple suppressive cues and significantly improved therapeutic performance across diverse solid tumor models in both CAR and TCR systems. Moreover, combinatorial knockout of P2RY8 and GNAS further enhanced overall tumor control, demonstrating that complementary screens with distinct in vivo readouts can nominate gene targets addressing orthogonal barriers, and that combining such edits can improve therapeutic potency. This flexible and scalable platform can be adapted to diverse tumor models and screening modalities, enabling systematic discovery of genetic strategies to equip T cell therapies for solid tumors.

ORGANISM(S): Homo sapiens

PROVIDER: GSE330227 | GEO | 2026/06/15

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Medulloblastoma, the most common malignant pediatric brain tumor, is highly heterogeneous with distinct molecular subtypes and cellular origins. Although current treatments improve survival rates, patients suffer severe treatment-related side effects and often relapse of tumors carrying resistance mutations, underscoring an urgent need for alternative targeted therapies. Currently, the genetic alterations underlying this disease are not fully understood. Here we identify GNAS, encoding the G-protein Gs-alpha, as a potent tumor suppressor gene in medulloblastoma. GNAS specifically defines a subset of aggressive Sonic Hedgehog (Shh)-group medulloblastomas. Gnas loss-of-function in distinct lineage progenitors of the developing hindbrain suffices to initiate medulloblastoma. We find that Gs-alpha is highly enriched at primary cilia of granule neuron precursors and suppresses Shh signaling not only by regulating classic cAMP-dependent pathway but also controlling ciliary trafficking of Smoothened. Concurrent cAMP elevation and Smoothened inhibition robustly arrests tumor cell growth in Gnas mutants. We further reveal oligodendrocyte progenitors as a novel cellular origin for anatomically-distinct Shh-associated medulloblastomas. Together, we identify a previously unrecognized tumor suppressor function of Gs-alpha in medulloblastoma partially mediated through inhibiting Shh signaling, and uncover Gs-alpha as a molecular link across disparate cells of origin among Shh-group medulloblastomas, pointing to G- protein modulation as a potential therapeutic avenue. Purpose: To known the gene expression profile of Medulloblastoma which drived by Gnas mutation Methods: mRNAs isolated from the cerebellum of control and Gnas mutants Results:Upregulation of Shh Signaling components in tumors Conclusions: The deletion of Gnas in hGFAP and Olig1 possitive cells result in substantial upregulation of shh signaling and formation of Medulloblastoma cerebellum mRNA profiles of 3 60-day old wild type (Ctrl) and 8 Olig1Cre driven Gsa conditional knockout or 8 hGFAPCre driven conditional knockout mice were generated by deep sequencing using Illumina Hiseq2000

Dataset Information

In vivo genome-wide CRISPR screens in human T cells to enhance T cell therapy for solid tumors

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets