Modulatable switch for selection of donor modified cells

Abstract

The disclosed methods are generally directed to preventing, treating, suppressing, controlling or otherwise mitigating side effects of T-cell therapy, the T-cell therapy designed to accelerate immune reconstitution, induce a GVM effect, and/or target tumor cells.

1 . A method of providing benefits of a lymphocyte infusion to a patient in need of treatment thereof comprising:
 (a) generating HPRT deficient lymphocytes from a donor sample, wherein the HPRT deficient lymphocytes are generated through knockout of the HPRT gene;   (b) positively selecting for the generated HPRT deficient lymphocytes ex vivo to provide a population of modified lymphocytes; and   (c) administering at least a portion of the population of modified lymphocytes to the patient following an administration of an HSC graft to the patient.   
     
     
         2 . The method of  claim 1 , further comprising administering one or more doses of methotrexate or mycophenolic acid if side effects from the administration of the at least the portion of the population of modified lymphocytes arises. 
     
     
         3 . The method of  claim 2 , wherein an amount of methotrexate administered ranges from about 2 mg/m 2 /infusion to about 100 mg/m 2 /infusion. 
     
     
         4 . The method of  claim 2 , wherein the methotrexate or mycophenolic acid is administered in titrated doses. 
     
     
         5 . The method of  claim 1 , wherein the positive selection comprises contacting the generated HPRT deficient lymphocytes with a purine analog. 
     
     
         6 . The method of  claim 5 , wherein the purine analog is selected from the group consisting of 6-thioguanine, 6-mercaptopurine, and azathiopurine. 
     
     
         7 . The method of  claim 6 , wherein the purine analog is 6-thioguanine. 
     
     
         8 . The method of  claim 1 , wherein the positive selection comprises contacting the generated HPRT deficient lymphocytes with both a purine analog and allopurinol. 
     
     
         9 . The method of  claim 1 , wherein the modified lymphocytes are administered as a single bolus. 
     
     
         10 . The method of  claim 1 , wherein multiple doses of the modified lymphocytes are administered. 
     
     
         11 . The method of  claim 10 , wherein each dose of the modified lymphocytes comprises between about 0.1×10 6  cells/kg to about 240×10 6  cells/kg. 
     
     
         12 . The method of  claim 1 , wherein the knockout of the HPRT gene comprises contacting lymphocytes with an HPRT-targeted CRISPR/Cas9 RNP. 
     
     
         13 . The method of  claim 12 , wherein the HPRT-targeted CRISPR/Cas9 RNP is formulated within a nanocapsule. 
     
     
         14 . The method of  claim 1 , wherein the knockout of the HPRT gene comprises contacting lymphocytes with CRISPR/Cas9 and an anti-HPRT sgRNA. 
     
     
         15 . The method of  claim 14 , wherein at least one of the Cas9 or the anti-HPRT sgRNA are expressed from a vector. 
     
     
         16 . The method of  claim 15 , wherein the vector is a non-integrating lentiviral vector or a non-reverse transcribed lentiviral vector. 
     
     
         17 . A method of treating a hematological cancer in a patient in need of treatment thereof comprising:
 (a) generating HPRT deficient lymphocytes from a donor sample, wherein the HPRT deficient lymphocytes are generated through knockout of the HPRT gene;   (b) positively selecting for the generated HPRT deficient lymphocytes ex vivo to provide a population of modified lymphocytes;   (c) inducing at least a partial graft versus malignancy effect by administering an HSC graft to the patient; and   (d) administering at least a portion of the population of modified lymphocytes to the patient following the detection of residual disease or disease recurrence.   
     
     
         18 . The method of  claim 17 , wherein the method further comprises administering one or more doses of methotrexate or mycophenolic aid. 
     
     
         19 . The method of  claim 18 , wherein an amount of methotrexate administered ranges from about 2 mg/m 2 /infusion to about 100 mg/m 2 /infusion. 
     
     
         20 . The method of  claim 18 , wherein the methotrexate or mycophenolic is administered in titrated doses. 
     
     
         21 . The method of  claim 17 , wherein the positive selection comprises contacting the generated HPRT deficient lymphocytes with a purine analog selected from the group consisting of 6-thioguanine, 6-mercaptopurine, and azathiopurine. 
     
     
         22 . The method of  claim 17 , wherein multiple doses of the modified lymphocytes are administered. 
     
     
         23 . The method of  claim 22 , wherein each dose of the modified lymphocytes comprises between about 0.1×10 6  cells/kg to about 240×10 6  cells/kg. 
     
     
         24 . The method of  claim 17 , wherein the knockout of the HPRT gene comprises contacting lymphocytes with CRISPR/Cas9 and an anti-HPRT sgRNA.