Tumor infiltrating lymphocytes and methods of therapy
Inventors
Moriarity, Branden • Webber, Beau • Choudhry, Modassir • Rosenberg, Steven A. • Palmer, Douglas C. • Restifo, Nicholas P.
Assignees
University of Minnesota Twin Cities • Intima Bioscience Inc • University of Minnesota System • US Department of Health and Human Services • Office of Technology Transfer
Publication Number
US-10912797-B2
Publication Date
2021-02-09
Expiration Date
2037-10-18
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Abstract
Genetically modified compositions, such as non-viral vectors and tumor infiltrating lymphocytes, for the treatment of gastrointestinal cancer are disclosed. Disclosed are methods of utilizing a CRISPR system to generate genetically modified compositions. Also disclosed are the methods of making and using the genetically modified compositions for the treatment of gastrointestinal cancer.
Core Innovation
The invention provides genetically modified compositions, including tumor infiltrating lymphocytes (TILs) and non-viral vectors, for the treatment of gastrointestinal cancer. It discloses methods for utilizing a CRISPR system to generate these genetically modified compositions, and methods for making and using them therapeutically for gastrointestinal cancer.
The invention addresses the challenge of effective TIL therapy in solid tumors other than metastatic melanoma. These challenges stem from suppressive tumor microenvironments, intrinsic impairments in T-cell receptor signaling and effector functions, and risks associated with systemic immune checkpoint inhibitors that cause autoimmune side-effects. Current genetic engineering approaches have mainly succeeded in hematologic tumors but are limited in solid tumors due to lack of specific tumor-expressed molecular targets. Advances in identifying tumor-specific mutations that elicit T-cell responses, for example via whole-exomic-sequencing, are harnessed in this invention to generate mutation-reactive TILs genetically engineered to disrupt at least a portion of the cytokine inducible SH2-containing protein (CISH) gene using CRISPR systems.
Claims Coverage
The patent contains multiple independent claims focusing on therapeutic methods and compositions involving tumor infiltrating lymphocytes with genomic disruption of the CISH gene, and administration protocols involving immunosuppressants, antifungals, antibiotics, and immunostimulants.
Use of tumor infiltrating lymphocytes with CISH gene disruption for cancer treatment
Administration to a subject of a pharmaceutical composition comprising tumor infiltrating lymphocytes (TILs) where CD4+ T cells have a genomic disruption of at least a portion of the cytokine inducible SH2-containing protein (CISH) gene that suppresses or eliminates CISH protein expression to treat cancer.
Preparative immunosuppressive regimen combined with antifungal and TIL therapy
A lymphodepleting preparative immunosuppressive regimen sufficient to cause lymphodepletion is administered along with a pharmaceutical composition comprising an antifungal agent inhibiting fungal infection and TILs with genomic disruption of the CISH gene for treating cancer.
Inclusion of antibiotic agents in the treatment method
The method further includes administering antibiotics sufficient to inhibit bacterial infections, including cephalosporins, quinolones, penicillins, rifamycins, and other classes specifically listed in the claims.
Administration of immunostimulants with TIL therapy
The method optionally includes administering immunostimulants such as interleukins (IL-2, IL-7, IL-12, IL-15, IL-21) or G-CSF concurrently or sequentially with TIL therapy.
Specific dosing ranges for immunosuppressants and antifungals
Cyclophosphamide is administered at 50 to 80 mg/kg, and fludarabine at 20 to 30 mg/m2. Fluconazole or other azoles are included as antifungal agents. The preparative immunosuppressive regimen can include two immunosuppressants administered sequentially or simultaneously.
Therapeutic products comprising TILs with CISH disruption and prophylactic agents
Compositions comprising a pharmaceutical dosage form including TILs with disruption of CISH gene, antifungals, immunosuppressants, antibiotics and immunostimulants formulated for therapeutic administration are disclosed.
Treatment of gastrointestinal and other cancers
Methods are directed to treating gastrointestinal cancer and include autologous or allogeneic TILs genetically modified by CRISPR disruption of the CISH gene to suppress CISH protein expression.
The claims cover the use of genetically modified TILs with disrupted CISH gene for cancer therapy in combination with preparative immunosuppressive regimens, antifungal and antibiotic prophylaxis, immunostimulant administration, and specify dosing and treatment protocols, compositions and methods of manufacture for these therapeutic modalities.
Stated Advantages
Genetic disruption of the CISH gene in tumor-infiltrating lymphocytes enhances functional avidity, proliferation, cytokine polyfunctionality, and durable regression of tumors.
Use of CRISPR system allows efficient and specific knockout of immune checkpoint genes reducing immunosuppressive effects without systemic autoimmune side-effects.
Preparative regimes combining immunosuppressants with antifungal and antibiotic prophylaxis reduce immune rejection and infection risk during TIL therapy.
The methods permit large-scale expansion and cryopreservation of knockout TIL products suitable for clinical administration with maintained viability and function.
Documented Applications
The primary application is the treatment of gastrointestinal cancer in human subjects by administering autologous tumor infiltrating lymphocytes genetically disrupted in the CISH gene by CRISPR technology combined with immunosuppressive preparative regimens and infection prophylaxis.
Methods are also described for adoptive cell transfer therapy of metastatic cancer refractory to standard treatments, including melanoma, lymphoma, breast cancer, and prostate cancer, using CISH knockout mutation-reactive TILs.
The therapy includes isolation of TILs from tumor tissue, identification of mutation reactive TIL by whole-exomic sequencing and antigen presentation assays, engineering of TIL by CRISPR knockout of CISH, expansion, and administration with immunosuppression and immunostimulation.
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