Intracellular genomic transplant and methods of therapy
Inventors
Moriarity, Branden • Webber, Beau • Choudhry, Modassir • Rosenberg, Steven A. • Palmer, Douglas C. • Restifo, Nicholas P.
Assignees
Intima Bioscience Inc • University of Minnesota System • US Department of Health and Human Services
Publication Number
US-11642375-B2
Publication Date
2023-05-09
Expiration Date
2036-07-29
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Abstract
Genetically modified compositions, such as non-viral vectors and T cells, for treating cancer are disclosed. Also disclosed are the methods of making and using the genetically modified compositions in treating cancer.
Core Innovation
Despite significant advances in cancer therapeutics, many tumor types remain resistant to chemotherapy, radiotherapy, or biotherapy, particularly in advanced stages where surgical options are limited. Genetic engineering of lymphocytes to recognize molecular targets on tumors has shown remarkable complete remissions but these successes are largely limited to hematologic tumors. Broader application to solid tumors is limited by the lack of identifiable tumor-specific molecules that can be targeted to mediate tumor destruction.
The invention discloses genetically modified compositions and methods for treating cancer by identifying cancer-specific T cell receptors (TCRs) that recognize unique immunogenic mutations in a patient's cancer. These TCR transgenes are inserted into T cells using non-viral methods like CRISPR, TALEN, transposon-based technologies, or Mega-TAL, enabling extended immunotherapy to diverse cancer types. Engineered cells comprising gene disruptions, such as checkpoint gene disruptions (e.g., PD-1), and non-virally integrated TCR sequences produce functional TCRs that recognize cancer neoantigens in the context of major histocompatibility complexes.
Methods include non-viral introduction of polynucleic acids encoding exogenous TCR sequences flanked by recombination arms complementary to genomic sequences, often immune checkpoint genes, and the use of homologous recombination enhancers to facilitate integration at double-stranded break regions created by nucleases such as CRISPR. Modifiers that reduce cellular toxicity from polynucleic acid introduction, like caspase inhibitors, increase cell viability. Engineered cells can be selected, expanded ex vivo or in vivo, and used autologously or allogeneically in combination therapies to treat cancer, infections, autoimmune disorders, or graft-versus-host disease.
Claims Coverage
The patent includes two main independent claims describing isolated genetically modified human cells with specific genomic disruptions and engineered receptor sequences. The inventive features center on the genomic modifications, integration methods, and the functional attributes of the engineered receptors.
Engineered human T cells with genomic disruption and exogenous receptor integration
An isolated genetically modified human T cell comprising a genomic disruption within a cytokine inducible SH2-containing protein (CISH) gene target sequence comprising an endonuclease-mediated indel, and either complementary DNA or mRNA encoding an exogenous functional T cell receptor or chimeric antigen receptor flanked by homology arms binding adjacent to disruptions in T Cell Receptor Alpha Constant gene, T Cell Receptor Beta Locus gene, or CISH gene sequences, where the exogenous receptor binds a neoantigen.
Methods and systems for genomic disruption and receptor integration
The use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems to perform genomic disruption within the CISH gene sequence and to integrate exogenous TCR or chimeric antigen receptor DNA or mRNA in a targeted fashion at the indicated gene loci, producing cells with enhanced cancer targeting capabilities.
Cancer targeting functional receptor expression
Expression of exogenous functional T cell receptors or chimeric antigen receptors in engineered human cells that bind antigens or neoantigens expressed by cancer cells across a wide spectrum of cancer types, with integrations adjacent to or within critical immune-related gene loci such as CISH, PD-1, and others.
The claims cover isolated genetically modified human cells, particularly T cells, with genomic disruptions at CISH gene sequences via endonuclease activity and integration of exogenous TCR or chimeric antigen receptor DNA or mRNA in a targeted fashion at the indicated gene loci, where the exogenous receptor binds cancer neoantigens. The invention encompasses the use of CRISPR systems for these modifications and the therapeutic targeting of cancer neoantigens by the engineered receptors.
Stated Advantages
Provides high efficiency gene transfer and expression of tumor-specific TCRs into T cells.
Increases cell survival rates by reducing cytotoxicity from exogenous nucleic acids during genetic modification.
Facilitates efficient introduction of double strand breaks and favors homology-directed repair over non-homologous end joining, increasing precision of gene integration.
Enables reliable expression of functional TCRs recognizing unique cancer neoantigens, extending immunotherapy to diverse cancer types including solid tumors.
Allows engineering of autologous or non-autologous T cells compatible with good manufacturing practices (GMP) for clinical therapy.
Documented Applications
Identification of cancer-specific T cell receptors (TCRs) recognizing unique immunogenic mutations in a patient's cancer for cellular therapy.
Treatment of various cancers including bladder cancer, epithelial cancer, bone cancer, brain tumor, breast cancer, gastrointestinal cancer, leukemia, liver cancer, lung cancer, lymphoma, myeloma, ovarian cancer, prostate cancer, sarcoma, stomach cancer, thyroid cancer, hematologic malignancies, and solid tumors.
Use of engineered T cells comprising functional exogenous receptors to treat cancer patients by adoptive cell transfer-based immunotherapy.
Engineering of tumor infiltrating lymphocytes (TILs) and peripheral blood lymphocytes (PBLs) for administration to subjects in need thereof.
Combination therapy comprising engineered T cells and other agents such as chemotherapy, radiation, immunosuppressive drugs, bone marrow transplantation, or immunotherapy for enhanced cancer treatment.
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