Autologous tumor vaccines and methods

Abstract

Autologous anti-cancer vaccines and methods of manufacture and treatment are provided, including expansion of individual patient-derived tumor cells in an immune-compromised animal(s) to attain, quantitatively and qualitatively, sufficient material for efficacious vaccine production and utilization, to elicit an immune response against micrometastases and/or recurrence in the individual patient following tumor excision.

What is claimed as new and desired to be protected by Letters Patent is: 
     
         1 . A method for making an injectable autologous, anti-neoplastic vaccine containing at least about 10 7  viable, non-tumorigenic tumor cells, comprising:
 excising a solid tumor from a cancer patient to obtain at least 95% of the patient's solid tumor tissue, wherein the excised tumor tissue has a weight of less than about 3.5 grams;   digesting the excised tumor tissue to obtain dissociated tumor cells;   transplanting the excised tumor cells into an immune-compromised animal;   propagating the tumor cells in the animal to obtain expanded tumor tissue;   harvesting the expanded tumor tissue from the animal to obtain harvested tumor cells, wherein the harvested tumor tissue has a weight of at least about 3.5 grams;   applying a dose of gamma radiation to the harvested tumor cells to render the cells non-tumorigenic; and   combining the non-tumorigenic tumor cells with a pharmaceutically acceptable carrier for injection to prepare an injectable dose containing at least about 10 7  tumor cells.   
     
     
         2 . A method according to  claim 1 , wherein the expanded cells have at least 85% sequence homology to the tumor cells of the patient's solid tumor. 
     
     
         3 . A method according to  claim 1 , wherein the injectable dose comprises tumor cells having at least 80% viability. 
     
     
         4 . A method according to  claim 1 , further digesting the excised tumor tissue with an enzyme to obtain dissociated tumor cells and mixing the dissociated tumor cells to form a homogeneous suspension. 
     
     
         5 . A method according to  claim 3 , further comprising digesting the harvested tumor tissue to obtain dissociated harvested tumor cells and mixing the dissociated harvested tumor cells to form a homogeneous suspension. 
     
     
         6 . A method according to  claim 1 , wherein the tumor cells transplanted into the animal comprise about 95-100% by weight of the patient's solid tumor. 
     
     
         7 . A method according to  claim 5 , wherein the tumor cells harvested from the animal comprise about 95-100% by weight of the expanded tumor tissue. 
     
     
         8 . A method according to  claim 1 , further comprising transplanting the harvested tumor tissue into a second immune-compromised animal, propagating the tumor tissue in the second animal, and harvesting the propagated tumor tissue from the second animal to prepare the injectable dose containing at least about 10 7  tumor cells. 
     
     
         9 . A method according to  claim 1 , where the animal is a rat. 
     
     
         10 . A method according to  claim 1 , where the animal is an athymic nude mouse which has received treatment to reduce Natural Killer Cell activity or has Severe Combined Immunodeficiency Disease (SCID). 
     
     
         11 . A method according to  claim 1 , where the transplanted tissue is harvested when it has a weight of about 3.5 to 4 g. 
     
     
         12 . The method of  claim 1 , further comprising treatment of the harvested tumor, before dissociation, with a disinfectant solution at a concentration and for a duration that provides anti-microbial activity while minimizing cytotoxicity. 
     
     
         14 . The method of  claim 1 , further comprising characterizing the antigenic profile of the harvested cells via RNA sequencing to ensure that heterogeneity to the patient's tumor has been preserved. 
     
     
         15 . The method of  claim 1 , further comprising applying a dose of gamma radiation to the harvested cells in an amount sufficient to inactivate microorganisms and tumorigenicity and preserve the viability of the cells, to obtain sterile, non-tumorigenic and immunogenic tumor cells. 
     
     
         16 . A method for eliciting an immune response to prevent the recurrence of metastases in a cancer patient in need, comprising:
 excising a solid tumor from a cancer patient to obtain at least about 95% of the patient's solid tumor tissue, wherein the excised tumor tissue has a weight of less than about 3.5 grams;   digesting the excised tumor tissue to obtain dissociated tumor cells;   transplanting the tumor cells into an immune-compromised animal;   propagating the tumor cells in the animal to obtain expanded tumor tissue;   harvesting the expanded tumor tissue from the animal, wherein the harvested tumor tissue has a weight of at least about 3.5 grams;   applying a dose of gamma radiation to the harvested tumor cells to render the cells non-tumorigenic;   combining the non-tumorigenic tumor cells with a pharmaceutically acceptable carrier for injection to prepare an injectable dose containing at least about 10 7  tumor cells having at least 85% sequence homology to the cells of the patient's tumor; and   administering to the patient said injectable dose to elicit an immunogenic response against recurrence of said cancer.   
     
     
         17 . A method according to  claim 16 , further comprising combining the harvested tumor cells with a pharmaceutically acceptable carrier for injection to prepare at least four injectable doses each containing at least about 10 7  dissociated tumor cells having at least 85% sequence homology to the patient's excised tumor; and
 administering each of said at least four doses in a treatment regime sufficient to elicit an immune response against recurrence of said cancer.   
     
     
         18 . A method according to  claim 17 , wherein the harvested tumor cells have at least 95% sequence homology to the cells of the patient's tumor. 
     
     
         19 . A method according to  claim 17 , further comprising administering an adjuvant or immune stimulator with one or more of said doses. 
     
     
         20 . A method according to  claim 16 , wherein the tumor transplanted into the animal comprises about 95-100% by weight of the excised solid tumor and the harvested tumor tissue comprises about 95-100% by weight of the expanded tumor tissue. 
     
     
         21 . A method according to  claim 18 , further comprising applying a dose of about 150,000-200,000 rads of gamma radiation to the tumor cells to render the tumor cells sterile and non-tumorigenic. 
     
     
         22 . A method according to  claim 16 , wherein said patient in need has stage III or earlier cancer. 
     
     
         23 . A method according to  claim 16 , wherein said patient in need has stage II or earlier cancer. 
     
     
         24 . A method according to  claim 16 , wherein said patient has cancer selected from the group consisting of colon, renal, melanoma, ovarian, or breast cancer. 
     
     
         25 . The method of  claim 16 , wherein the said patient has colon cancer and said excising is performed during colonoscopy. 
     
     
         26 . The method of  claim 25 , further comprising:
 washing the excised tumor tissue with a wash solution containing a detergent;   treating the excised tumor tissue with a disinfectant to reduce microbial contamination of the tissue; and   digesting the tumor tissue with a dissociation enzyme, in the presence of at least one antibiotic and an anti-mycotic, to obtain at least about 10 7  dissociated tumor cells.   
     
     
         27 . A whole cell autologous anti-cancer vaccine composition for administration to a patient after surgery to excise a solid tumor having a mass less than about 3.5 g, said composition comprising:
 at least about 10 7  viable non-tumorigenic tumor cells having at least 85% sequence homology to the cells of the excised tumor; and   a pharmaceutically acceptable carrier for injection;   wherein said composition gives rise to an immunogenic response when administered via intradermal injection to said patient.   
     
     
         28 . A composition according to  claim 27 , wherein the composition comprises at least four doses each containing at least 10 7  viable non-tumorigenic tumor cells having at least 85% sequence homology to the cells of the excised tumor. 
     
     
         29 . A composition according to  claim 27 , wherein the vaccine comprises at least 10 8  viable non-tumorigenic tumor cells having at least 85% sequence homology to the cells of the excised tumor. 
     
     
         30 . A composition according to  claim 27 , wherein the tumor cells of the vaccine have at least 95% sequence homology to the patient's primary tumor cells. 
     
     
         31 . A composition according to  claim 27  wherein the viable tumor cells of the vaccine are sterile, non-tumorigenic, and at least 80% viable. 
     
     
         32 . A method for making an injectable, autologous, anti-neoplastic vaccine containing at least about 10 7  viable, non-tumorigenic tumor cells, comprising:
 excising a solid tumor from a cancer patient to obtain at least 95% of the patient's solid tumor tissue, wherein the excised tumor tissue has a weight of greater than 3.5 grams;   digesting all of the excised tumor tissue with an enzyme to obtain dissociated tumor cells;   mixing the dissociated tumor cells to prepare a homogeneous suspension of heterogeneous cells, wherein any aliquot of the suspension contains a full complement of antigenic material from the patient's solid tumor;   applying a dose of gamma radiation to the homogeneous suspension of tumor cells to render the cells non-tumorigenic; and   combining the non-tumorigenic tumor cells with a pharmaceutically acceptable carrier for injection to prepare an injectable dose containing at least about 10 7  tumor cells.   
     
     
         33 . A method according to  claim 32 , wherein non-tumorigenic cells in each dose have at least 98% sequence homology to the patient's tumor cells. 
     
     
         34 . A method according to  claim 32 , further comprising characterizing the antigenic profile of the cells to ensure original tumor heterogeneity has been preserved. 
     
     
         35 . A method according to  claim 32 , further comprising applying a dose of gamma radiation to the cells in an amount sufficient to inactivate microorganisms and tumorigenicity and preserve the viability of the cells, to obtain sterile, non-tumorigenic and immunogenic tumor cells. 
     
     
         36 . A method for eliciting an immune response to prevent the recurrence of metastases in a cancer patient in need, comprising administering to the patient an injectable dose according to  claim 32  to elicit an immunogenic response against recurrence of said cancer.