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

Patent metadata

Field	Value
Publication number	US-10350245-B2
Application number	US-201615545555-A
Country	US
Kind code	B2
Filing date	Jan 21, 2016
Priority date	Jan 21, 2015
Publication date	Jul 16, 2019
Grant date	Jul 16, 2019

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.

First claim

What is claimed is:

1. A method performed by a device, the method comprising:
receiving, by a treatment chamber, a subject sample
transferring, by at least one valve and at least one pump, the subject sample from the treatment chamber to at least one target cell selector;
separating, by the at least one target cell selector, target cells from non-target cells in the subject sample by allowing the non-target cells to pass into a waste bag and retaining the target cells within the at least one target cell selector;
transferring, by the at least one valve and the at least one pump, the target cells into the treatment chamber;
introducing a genetic modifier to the target cells to generate genetically-modified target cells;
pelleting the genetically-modified target cells within the treatment chamber through centrifugation to create a pelleted cell suspension;
removing a specified supernatant volume from the pelleted cell suspension in the treatment chamber through a first tubing set connected to the treatment chamber;
diluting the pelleted cell suspension with final formulation media comprising a pharmaceutically acceptable carrier;
centrifuging the diluted pelleted cell suspension and removing an additional volume of supernatant in the treatment chamber through the first tubing set to form a final cell product formulation; and
transferring the final cell product formulation into one or more cryobags through a second tubing set connected to the treatment chamber wherein the final cell product formulation is ready for administration to a subject upon completion of transfer into the one or more cryobags.

2. The method of claim 1 wherein a user of the device:
determines an initial volume of the subject sample;
determines a hematocrit level of the subject sample;
determines a volume of buffer to add to the subject sample to reduce the hematocrit level to at least 25%
according to the following formula:

(

starting
⁢

⁢
product
⁢

⁢
volume
⁢

⁢

(
mL
)

×
obtained
⁢

⁢
hematocrit
⁢

⁢
value
⁢

⁢

(
%
)

25
⁢
%
⁢

⁢
desired
⁢

⁢
hematocrit
⁢

⁢
value

)

-

starting
⁢

⁢
product
⁢

⁢
volume
⁢

⁢

(
mL
)

;

and wherein the device adds, by the at least one valve and the at least one pump, the determined volume of buffer to the subject sample.

3. The method of claim 1 comprising performing release testing to verify compliance with Current Good Manufacturing Practices wherein the release testing comprises

Test
Required Result

Gram Stain
Negative

 3 Day Sterility
Negative

14 Day Sterility
Negative

Mycoplasma
Negative

Endotoxin
≤0.5 EU/ml

Cell Viability by Trypan Blue Dye Exclusion
≥70%.

4. The method of claim 1 wherein the introducing of the genetic modifier inserts or alters a gene selected from ABCD1, ABCA3, ABLI, ADA, AKT1, APC, APP, ARSA, ARSB, BCL11A, BLC1, BLC6, BRCA1, BRCA2, BRIP1, C9ORF72, C46, 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, at least one of RPL3 through RPL40, RPLPO, RPLP1, RPLP2, at least one of RPS2 through RPS30, RPSA, SFTPB, SFTPC, SGSH, SLX4, SNCA, SOD1, SOX2, TERC, TERT, TDP43, TINF2, TK, ubiquilin 2, VHL, WAS and WT-I.

5. The method of claim 1 wherein the genetically-modified target cells are 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, and/or bone marrow fibroblasts.

6. The method of claim 1 wherein the genetically-modified target cells are CD34+HSPC.

7. The method of claim 1 wher

Assignees

• Hutchinson Fred Cancer Res

Inventors

• Adair Jennifer E
• Kiem Hans-Peter

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61K35/28 Primary

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• C12N15/8673

  Special methods for packaging systems · CPC title

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12Q1/68

  involving nucleic acids · CPC title