Point-of-care and/or portable platform for gene therapy

Abstract

A platform for ex vivo isolation, production, and formulation of genetically-modified cells is described. The platform utilizes a software-enabled point-of-care and/or portable device making gene therapy more widely available.

1 .- 80 . (canceled) 
     
     
         81 . A method performed by a device, the method comprising:
 receiving, by a chamber, cells and a genetic modifier wherein the receiving results in genetic modification of the cells to produce genetically-modified cells within the chamber,   centrifuging the genetically-modified cells within the chamber to create a pelleted cell suspension;   removing, from the chamber, a supernatant volume from the pelleted cell suspension;   diluting the pelleted cell suspension with a media comprising a pharmaceutically acceptable carrier within the chamber to create a diluted pelleted cell suspension;   centrifuging the diluted pelleted cell suspension within the chamber and removing an additional volume of supernatant to form a cell product formulation; and   transferring the cell product formulation into a sterile receptacle through a conduit connected to the chamber.   
     
     
         82 . The method of  claim 81 , wherein the sterile receptacle is sterilely welded to the conduit. 
     
     
         83 . The method of  claim 81 , further comprising administering the transferred cell product formulation to a subject from the sterile receptacle. 
     
     
         84 . The method of  claim 81 , wherein the cells comprise hematopoietic stem cells (HSC), hematopoietic progenitor cells (HPC), hematopoietic stem and progenitor cells (HSPC), T cells, natural killer cells, B cells, macrophages, monocytes, mesenchymal stem cells (MSC), white blood cells (WBC), mononuclear cells (MNC), endothelial cells (EC), stromal cells, or bone marrow fibroblasts. 
     
     
         85 . The method of  claim 81 , wherein the cells comprise CD34+ HSPC. 
     
     
         86 . The method of  claim 81 , wherein the genetic modifier comprises a non-integrating vector. 
     
     
         87 . The method of  claim 81 , wherein the genetic modifier comprises a viral vector. 
     
     
         88 . The method of  claim 87 , wherein the viral vector comprises a lentiviral vector, a gammaretroviral vector, a foamy viral vector, or an alpharetroviral vector. 
     
     
         89 . The method of  claim 87 , wherein the viral vector comprises a lentiviral vector comprising a pseudotype envelope glycoprotein and a lentiviral RNA molecule, wherein the pseudotype envelope glycoprotein comprises VSVG, cocal, RD114, or mFoamy, and wherein the lentiviral RNA molecule comprises an integration deficient HIV-1-derived, self-inactivating lentivirus backbone. 
     
     
         90 . The method of  claim 87 , wherein the viral vector comprises a foamy viral vector comprising a pseudotype envelope glycoprotein and a foamy viral RNA molecule wherein the pseudotype envelope glycoprotein comprises Foamy, or mFoamy and wherein the foamy viral RNA molecule comprises an integration deficient self-inactivating foamy virus backbone. 
     
     
         91 . The method of  claim 81 , wherein the genetic modifier inserts or alters a gene within the cells. 
     
     
         92 . The method of  claim 91 , wherein the gene comprises ABCD1, ABCA3, ABLI, ADA, AKT1, APC, APP, ARSA, ARSB, BCL11A, BLC1, BLC6, BRCA1, BRCA2, BRIP1, C9ORF72, C46 or other C peptide, CAR, CAS9, C-CAM, CBFAI, CBL, CCR5, CD4, CD19, CD40, CDA, CFTR, CLN3, C-MYC, CRE, CSCR4, CSFIR, CTLA, CTS-I, CYB5R3, DCC, DHFR, DKC1, DLL1, DMD, EGFR, ERBA, ERBB, EBRB2, ETSI, ETS2, ETV6, F8, F9, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, FasL, FCC, FGR, FOX, FUS, FUSI, FYN, GALNS, GATA1, GLB1, GNS, GUSB, HBB, HBD, HBE1, HBG1, HBG2, HCR, HGSNAT, HOXB4, HRAS, HYAL1, ICAM-1, iCaspase, IDUA, IDS, JUN, KLF4, KRAS, LCK, LRRK2, LYN, MCC, MDM2, MGMT, MLL, MMACI, MYB, MEN-I, MEN-II, MYC, NAGLU, NANOG, NF-1, NF-2, NKX2.1, NOTCH, OCT4, p16, p21, p27, p53, p57, p73, PALB2, PARK2, PARK7, phox, PINK1, PK, PSEN1, PSEN2, PTPN22, RAD51C, ras, RPL3 through RPL40, RPLP0, RPLP1, RPLP2, RPS2 through RPS30, RPSA, SFTPB, SFTPC, SGSH, SLX4, SNCA, SOD1, SOX2, TERC, TERT, TDP43, TINF2, TK, ubiquilin 2, VHL, WAS or WT-1. 
     
     
         93 . The method of  claim 81 , further comprising receiving, by the chamber, transduction media comprising a base medium and a cytokine comprising human granulocyte colony stimulating factor, stem cell factor, thrombopoietin, flightless 3 ligand, interleukin 3 and/or interleukin 6. 
     
     
         94 . The method of  claim 81 , wherein the chamber receives the cells in a number of stages. 
     
     
         95 . A method of preparing a genetically-modified cell formulation, the method performed by a closed-system device, the method comprising:
 receiving, by a chamber,
 a subject sample comprising cells; 
 a pharmaceutically acceptable carrier; and 
 a genetic modifier; 
   removing, from the chamber, a first volume of the pharmaceutically acceptable carrier through a conduit connected to the chamber;   receiving, by the chamber, a second volume of the pharmaceutically acceptable carrier through a conduit connected to the chamber thereby forming the genetically-modified cell formulation; and   transferring the genetically-modified cell formulation into one or more sterile receptacles through at least one conduit connected to the chamber wherein no further cell processing is required before administering the final genetically-modified cell formulation to a subject.   
     
     
         96 . The method of  claim 95 , wherein the sterile receptacle is sterilely welded to the conduit. 
     
     
         97 . The method of  claim 95 , further comprising administering the transferred cell product formulation to a subject from the sterile receptacle. 
     
     
         98 . The method of  claim 95 , wherein the cells comprise hematopoietic stem cells (HSC), hematopoietic progenitor cells (HPC), hematopoietic stem and progenitor cells (HSPC), T cells, natural killer cells, B cells, macrophages, monocytes, mesenchymal stem cells (MSC), white blood cells (WBC), mononuclear cells (MNC), endothelial cells (EC), stromal cells, or bone marrow fibroblasts. 
     
     
         99 . The method of  claim 95 , wherein the genetic modifier comprises a non-integrating viral vector. 
     
     
         100 . The method of  claim 95 , wherein the genetic modifier inserts or alters a gene within the cells.