Cryopreservation of apoptotic cancer cells for use in immunotherapy against cancer
Inventors
Fucikova, Jitka • Koci, Lenka • Pokorna, Katerina • Truxova, Iva • Moserova, Irena • Rozkova, Daniela • Spisek, Radek
Assignees
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Publication Number
US-9895430-B2
Publication Date
2018-02-20
Expiration Date
Abstract
Described herein is a reliable method for preparing a potent vaccine useful for immunotherapy comprising the step of cryopreserving a population of cells undergoing immunogenic cell death, and using such cells to activate dendritic cells for use in immunotherapy. In a specific embodiment, the method comprises cryopreserving cancer cells undergoing cell death, which can be used to prepare a pharmaceutical composition for immunotherapy against cancer.
Core Innovation
The disclosed subject matter relates to a method for preparing a pharmaceutical composition for use in immunotherapy. The method includes inducing immunogenic cell death in a population of cancer cells, and cryopreserving the cancer cells undergoing immunogenic cell death in a cryopreservant within six hours after induction of immunogenic cell death.
After thawing, the method includes putting the cancer cells back into culture and pulsing immature dendritic cells with the thawed cancer cells. The thawed cancer cells retain one or more of the hallmarks of immunogenic cell death, and maintaining immunogenic cell death-associated hallmarks during cryopreservation and subsequent handling is positioned as a functional requirement for preparing the immunotherapy composition.
The dependent claim refinements further identify specific hallmarks to be retained, including calreticulin and/or HSP70 and/or HSP90. Additional refinements specify cryopreservant selection and concentration, such as dimethyl sulphoxide at a concentration of at least 5%, a cryopreservation temperature below -75°C, a thawing time and temperature window, and a solid tumor cancer selected from prostate cancer, ovarian cancer, lung cancer, renal cancer, colon cancer, breast cancer, or glioblastoma.
Claims Coverage
The consolidated content includes one independent claim defining the full immunotherapy-pharmaceutical preparation workflow, supported by dependent claims that refine cryopreservant, time/temperature windows, hallmark selection, and cancer target scope. The inventive features focus on preserving immunogenic cell death hallmarks through cryopreservation and subsequent use in dendritic cell pulsing.
Immunogenic cell death in cancer cells with a post-induction cryopreservation window
A method for preparing a pharmaceutical composition for use in immunotherapy comprising inducing immunogenic cell death in a population of cancer cells, and cryopreserving the cancer cells undergoing immunogenic cell death in a cryopreservant, wherein the cells undergoing immunogenic cell death are cryopreserved within six hours after induction of immunogenic cell death.
Thawing and returning cryopreserved ICD-stage cells to culture
Thawing the cryopreserved cancer cells, wherein the thawed cells are put back into culture.
Pulsing immature dendritic cells with thawed cells retaining ICD hallmarks
Pulsing immature dendritic cells with the thawed cancer cells, wherein the thawed cancer cells retain one or more of the hallmarks of immunogenic cell death.
Cryopreservant selection for ICD-stage cell cryopreservation
The method further includes cryopreserving the cells undergoing immunogenic cell death in a cryopreservant containing dimethyl sulphoxide at a concentration of at least 5%.
Cryopreservation temperature constraint for ICD-stage cells
The method further includes cryopreserving the cells that undergo immunogenic cell death at a temperature below -75°C.
Thawing time and temperature window for ICD-stage cells
The thawing step is carried out for between 5 seconds and 10 minutes at a temperature between 32°C and 40°C.
Retention of selected immunogenic cell death hallmarks
The method further specifies that the selected hallmarks of immunogenic cell death include calreticulin, HSP70, and/or HSP90.
Solid tumor cancer restriction to enumerated tumor types
The method is applied to a solid tumor cancer selected from prostate cancer, ovarian cancer, lung cancer, renal cancer, colon cancer, breast cancer, or glioblastoma.
Overall, the claim coverage centers on preparing an immunotherapy pharmaceutical composition by cryopreserving cancer cells after inducing immunogenic cell death within a defined post-induction window, thawing and returning the cells to culture, and pulsing immature dendritic cells with the thawed cells while requiring retention of immunogenic cell death hallmarks. Dependent features further narrow cryopreservant composition, cryopreservation temperature, thawing time/temperature, which hallmarks must be retained, and the solid tumor cancer types.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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