IP Library › Granted Patent US 12,275,963
Granted Patent B2
US 12,275,963 · App. 16/611,383 · Granted Apr 15, 2025

Artificially manipulated immune cell

Inventors: Seok Joong Kim (Seoul, KR); Yoon-Young Kim (Seoul, KR); Ho-Sung Yu (Gyeonggi-do, KR); In-Young Jung (Gyeonggi-do, KR); Jung Min Lee (Gyeongsangbuk-do, KR)
Assignee: TOOLGEN INCORPORATED
C12N9/22A61K39/4611A61K39/4631A61K39/4644C07K14/705C07K16/2863C12N5/0634C12N5/0636C12N15/102C12N15/113C12N15/115C12N15/117C12N15/87C12Y207/01107A61K2239/31A61K2239/38C07K14/7051C07K2317/622C07K2319/03C12N2501/515
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,275,963
App. No.
16/611,383
Granted
Apr 15, 2025
Kind
B2
Abstract

The presents invention relates to a composition for manipulating an immune cell which is used for artificially manipulating an immune cell. More particularly, the present invention relates to a composition for manipulating an immune cell which is used for artificially manipulating an immune cell and a manipulated immune cell comprising an artificially modified immunity regulating gene and an artificial receptor which is produced using the composition, and use thereof.

Claims (27)

1. A manipulated human T cell with an enhanced immune activity comprising:

an engineered genome comprising an engineered endogenous DGKA gene and an engineered endogenous DGKZ gene; and

at least one artificial receptor and/or nucleic acid encoding the artificial receptor,

wherein the engineered endogenous DGKA gene comprises a first artificial modification,

wherein the first artificial modification is a first indel by CRISPR/Cas gene editing and induced in a region of exon 7 of the wild-type DGKA gene, wherein the engineered endogenous DGKA gene does not comprise any further modification other than the first artificial modification,

wherein the engineered endogenous DGKZ gene comprises a second artificial modification,

wherein the second artificial modification is a second indel by CRISPR/Cas gene editing and induced in a region of exon 3 of the wild-type DGKZ gene, wherein the engineered endogenous DGKZ gene does not comprise any further modification other than the second artificial modification,

wherein the first artificial modification causes at least one of a reduced expression of engineered endogenous DGKA gene and a functional impairment of the protein expressed from the engineered endogenous DGKA gene, and the second artificial modification causes at least one of a reduced expression of engineered endogenous DGKZ gene and a functional impairment of the protein expressed from the engineered endogenous DGKZ gene, in a manner that the manipulated human T cell has the enhanced immune activity.

2. The manipulated human T cell according to claim 1 , wherein the artificial receptor has a binding specificity for at least one antigen selected from the group consisting of A33, ALK, alpha-fetoprotein (AFP), adrenoreceptor beta 3 (ADRB3), alpha-folate receptor, AD034, AKT1, BCMA, beta-human chorionic gonadotropin, B7H3 (CD276), BST2, BRAP, CD5, CD13, CD19, CD20, CD22, CD24, CD30, CD33, CD38, CD40, CD44v6, CD52, CD72, CD79a, CD79b, CD89, CD97, CD123, CD138, CD160, CD171, CD179a, carbonic anhydrase IX (CAIX), CA-125, carcinoembryonic antigen (CEA), CCR4, C-type lectin-like molecules (CLL-1 or CLECL1), claudin6 (CLDN6), CXORF61, CAGE, CDX2, CLP, CT-7, CT8/HOM-TES-85, cTAGE-1, ERBB2, epidermal growth factor receptor (EGFR), EGFR variants III (EGFRvIII), epithelial cell adhesion molecule (EPCAM), E74-like factor 2 mutation (ELF2M), Ephrin type-A receptor 2 (EphA2), EMR2, Fms-like tyrosine kinase 3 (FLT3), FCRL5, fibulin-1, G250, GD2, glycoprotein 36 (gp36), glycoprotein 100 (gp100), glucocorticoid-induced tumor necrosis factor receptor (GITR), GPRC5D, GloboH, G protein-coupled receptor 20 (GPR20), GPC3, hsp70-2, human high molecular weight-melanoma-associated antigen (HMWMAA), hepatitis A virus cellular receptor 1 (HAVCR1), human papillomavirus E6 (HPV E6), human papillomavirus E7 (HPV E7), HAGE, HCA587/MAGE-C2, hCAP-G, HCE661, HER2/neu, HLA-Cw, HOM-HD-21/galectin9, HOM-MEEL-40/SSX2, HOM-RCC-3.1.3/CAXII, HOXA7, HOXB6, Hu, HUB 1, insulin growth factor (IGF1)-I, IGF-II, IGFI receptor, interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2), interleukin 11 receptor alpha (IL-11Ra), IGLL1, KIT (CD117), KM-HN-3, KM-KN-1, KOC1, KOC2, KOC3, KOC3, LAGA-1a, LAGE-1, LAIR1, LILRA2, LY75, Lewis Y antigen, MUC1, MN-CA IX, M-CSF, MAGE-1, MAGE-4a, mesothelin, MAGE-A1, MAD-CT-1, MAD-CT-2, MART1, MPPI 1, MSLN, neural cell adhesion molecule (NCAM), NY-ESO-1, NY-ESO-5, Nkp30, NKG2D, NY-BR-1, NY-BR-62, NY-BR-85, NY-CO-37, NY-CO-38, NNP-1, NY-LU-12, NY-REN-10, NY-REN-19/LKB/STK1 1, NY-REN-21, NY-REN-26/BCR, NY-REN-3/NY-CO-38, NY-REN-33/SNC6, NY-REN-43, NY-REN-65, NY-REN-9, NY-SAR-35, o-acetyl-GD2 ganglioside (OAcGD2), OGFr, PSMA, prostatic acid phosphatase (PAP), p53, prostate carcinoma tumor antigen-1 (PCTA-1), prostate stem cell antigen (PSCA), serine protease 21 (testisin or PRSS21), platelet-derived growth factor receptor-beta (PDGFR-beta), PLAC1, pannexin 3 (PANX3), PLU-1, ROR-1, RAGE-1, RU1, RU2, Rab38, RBPJ kappa, RHAMM, stage-specific embryonic antigen-4 (SSEA-4), SCP1, SSX3, SSX4, SSX5, Tyrp-1, TAG72, thyroglobulin, human telomerase reverse transcriptase (hTERT), 5T4, tumor-associated glycoprotein (TAG72), tyrosinase, transglutaminase 5 (TGS5), TEM1, TEM7R, thyroid-stimulating hormone receptor (TSHR), Tie 2, TRP-2, TOP2A, TOP2B, uroplakin 2 (UPK2), vimentin, vascular endothelial growth factor receptor 2 (VEGFR2), Wilms tumor protein 1 (WT1), and lewis (Y) antigen.

3. The manipulated human T cell according to claim 1 , wherein the artificial receptor is a chimeric antigen receptor (CAR).

4. The manipulated human T cell according to claim 1 , wherein the first artificial modification is induced in a first sequence constituting 50 nucleotides comprising a sequence selected from SEQ ID NOs: 20 to 24 of the wild-type DGKA gene, thereby the engineered endogenous DGKA gene does not comprise a sequence same as the first sequence.

5. The manipulated human T cell according to claim 1 , wherein the first artificial modification is induced in a first sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 23 of the wild-type DGKA gene, thereby the engineered endogenous DGKA gene does not comprise a sequence same as the first sequence.

6. The manipulated human T cell according to claim 1 , wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence selected from SEQ ID NOs: 109 to 113 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

7. The manipulated human T cell according to claim 1 , wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 111 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

8. The manipulated human T cell according to claim 1 , wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 113 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

9. The manipulated human T cell according to claim 1 , wherein the first artificial modification is induced in a first sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 23 of the wild-type DGKA gene, thereby the engineered endogenous DGKA gene does not comprise a sequence same as the first sequence, wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence selected from SEQ ID NOs: 109 to 125 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

10. The manipulated human T cell according to claim 1 , wherein the first artificial modification is induced in a first sequence constituting 50 nucleotides comprising a sequence selected from SEQ ID NOs: 19 to 24 of the wild-type DGKA gene, thereby the engineered endogenous DGKA gene does not comprise a sequence same as the first sequence, wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence selected from SEQ ID NOs: 109 to 113, 116, 120, 121, and 123 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

11. The manipulated human T cell according to claim 1 , wherein the first artificial modification is induced in a first sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 23 of the wild-type DGKA gene, thereby the engineered endogenous DGKA gene does not comprise a sequence same as the first sequence, wherein the second artificial modification is induced in a second sequence constituting 50 nucleotides comprising a sequence of SEQ ID NO: 113 of the wild-type DGKZ gene, thereby the engineered endogenous DGKZ gene does not comprise a sequence same as the second sequence.

12. A composition for immunotherapy comprising a manipulated human T cell with an enhanced immune activity, wherein the manipulated human T cell comprises:

an engineered genome comprising an engineered endogenous DGKA gene and engineered endogenous DGKZ gene; and

at least one artificial receptor and/or nucleic acid encoding the artificial receptor,

wherein the engineered endogenous DGKA gene comprises a first artificial modification,

wherein the first artificial modification is a first indel by CRISPR/Cas gene editing and induced in a region of exon 7 of the wild-type DGKA gene, wherein the engineered endogenous DGKA gene does not comprise any further modification other than the first artificial modification,

wherein the engineered endogenous DGKZ gene comprises a second artificial modification,

wherein the second artificial modification is a second indel and induced in a region of exon 3 of the wild-type DGKZ gene, wherein the engineered endogenous DGKZ gene does not comprise any further modification other than the second artificial modification,

wherein the first artificial modification causes at least one of a reduced expression of engineered endogenous DGKA gene and a functional impairment of the protein expressed from the engineered endogenous DGKA gene, and the second artificial modification causes at least one of a reduced expression of engineered endogenous DGKZ gene and a functional impairment of the protein expressed from the engineered endogenous DGKZ gene, in a manner that the manipulated human T cell has the enhanced immune activity.

13. The manipulated human T cells according to claim 1 , wherein the nucleic acid encoding the artificial receptor is in the engineered genome.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 6, 2019
From: KIM, SEOK JOONG; KIM, YOON-YOUNG; YU, HO-SUNG; JUNG, IN-YOUNG; LEE, JUNG MIN
To: TOOLGEN INCORPORATED
Reel/Frame 050934/0436 →
Priority Claims (1)
WO PCT/KR2017/008835 · Aug 14, 2017 · international
Continuity (3)
Provisional Application 62595159 · Dec 6, 2017
Provisional Application 62502822 · May 8, 2017
Related Publication 20210147798A1 · May 20, 2021
References Cited (93)
US 8207316B1 · Bentwich · 2012 [cited by examiner]
US 8865406B2 · Zhang · 2014 [cited by examiner]
US 10876120B2 · Wucherpfennig · 2020 [cited by examiner]
US 11041173B2 · Zhang et al. · 2021 [cited by applicant]
US 20050112568A1 · Friedman · 2005 [cited by examiner]
US 20050221354A1 · Mounts · 2005 [cited by applicant]
US 20050244851A1 · Blume et al. · 2005 [cited by applicant]
US 20110166037A1 · Cao et al. · 2011 [cited by applicant]
US 20130129668A1 · Firestein et al. · 2013 [cited by applicant]
US 20140120622A1 · Gregory et al. · 2014 [cited by applicant]
US 20150224142A1 · Albelda et al. · 2015 [cited by applicant]
US 20160120906A1 · Galetto et al. · 2016 [cited by applicant]
US 20160184362A1 · Duchateau et al. · 2016 [cited by applicant]
US 20160272999A1 · Duchateau et al. · 2016 [cited by applicant]
US 20170204372A1 · Mohler et al. · 2017 [cited by applicant]
US 20170335281A1 · Loew et al. · 2017 [cited by applicant]
US 20180119140A1 · Porteus et al. · 2018 [cited by applicant]
US 20190185860A1 · Kim et al. · 2019 [cited by applicant]
US 20190388468A1 · Lock · 2019 [cited by examiner]
US 20200299686A1 · Kwong · 2020 [cited by examiner]
US 20210128616A1 · Dave · 2021 [cited by examiner]
US 20210147798A1 · Kim et al. · 2021 [cited by applicant]
US 20210317406A1 · Marson · 2021 [cited by examiner]
AU 2014366047A1 · 2016 [cited by applicant]
AU 2013359123B2 · 2016 [cited by applicant]
CN 105121648A · 2015 [cited by applicant]
CN 105164264A · 2015 [cited by applicant]
EP 3498846A1 · 2019 [cited by applicant]
JP 2017500869A · 2017 [cited by applicant]
JP 2019524140A · 2019 [cited by applicant]
KR 1020150016588A · 2015 [cited by applicant]
KR 1020150105635A · 2015 [cited by applicant]
KR 1020160018425A · 2016 [cited by applicant]
KR 1020160138404A · 2016 [cited by applicant]
KR 1020170032406A · 2017 [cited by applicant]
WO WO2013176772A1 · 2013 [cited by applicant]
WO WO2014039513A2 · 2014 [cited by applicant]
WO WO2014186585A2 · 2014 [cited by applicant]
WO WO2015090230A1 · 2015 [cited by applicant]
WO WO2015121454A1 · 2015 [cited by applicant]
WO WO2016021972A1 · 2016 [cited by applicant]
WO WO2016069283A1 · 2016 [cited by applicant]
WO WO2016080097A1 · 2016 [cited by applicant]
WO WO2016123578A1 · 2016 [cited by applicant]
WO WO2018030874A1 · 2018 [cited by applicant]
Prinz, P. U., “High DGK- and Disabled MAPK Pathways Cause Dysfunction of Human Tumor-Infiltrating CD8+ T Cells That Is Reversible by Pharmacologic Intervention”, [cited by applicant]
Riese, M. J., et al.; “Enhanced Effector Responses in Activated CD8♭ T Cells Deficient in Diacylglycerol Kinases”, Cancer Res; 73(12) Jun. 15, 2013, pp. 3566-3577. [cited by applicant]
Cencic, R. et al.; “Protospacer Adjacent Motif (PAM)-Distal Sequences Engage CRISPR Cas9 DNA Target Cleavage”, PLOS One, Oct. 2014, vol. 9, Issue 10, pp. 1-13. [cited by applicant]
Riese, M. J., et al.; “Diacylglycerol Kinases (DGKs): Novel Targets for Improving T Cell Activity in Cancer”, Frontiers in Cell and Developmental Biology, Oct. 2016, vol. 4, Article 108, pp. 1-7. [cited by applicant]
Jung, I. Y., et al.; “CRISPR/Cas9-Medicated Knockout of DGK Improves Antitumor Activities of Human T Cells”, Cancer Res., 78(16), Aug. 15, 2018, pp. 4692-4703. [cited by applicant]
Extended European Search Report from corresponding European Patent Application No. 17839891.3, dated Feb. 11, 2020. [cited by applicant]
Search Report and Written Opinion from corresponding Singapore Patent Application No. 11201901184Q, dated Jun. 1, 2020. [cited by applicant]
Su S., et al.; “CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients”, Sci Rep., Jan. 28, 2016, vol. 6:20070, pp. 1-13. [cited by applicant]
1st Office Action from corresponding Australian Patent Application No. 2017308473, dated Aug. 14, 2020. [cited by applicant]
2nd Office Action from corresponding Australian Patent Application No. 2017308473, dated Sep. 11, 2020. [cited by applicant]
Examination Report from corresponding Russian Patent Application No. 2019106669, dated Sep. 23, 2020. [cited by applicant]
Office Action from corresponding European Patent Application No. 17839891.3, dated Oct. 23, 2020. [cited by applicant]
Office Action from corresponding Korean Patent Application No. 10-2019-0068999, dated Sep. 17, 2020. [cited by applicant]
Prinz et al., “NK?cell dysfunction in human renal carcinoma reveals diacylglycerol kinase as key regulator and target for therapeutic intervention” International Jornal of Cancer, vol. 135, No. 8, pp. 1832-1841, 2014. [cited by applicant]
Yang et al, “Diacylglycerol Kinase z Is a Target To Enhance NK Cell Function”, The Juornal of Immunology, vol. 197, No. 3, pp. 934-941, 2016. [cited by applicant]
Office Action from corresponding U.S. Appl. No. 16/324,955, issued on Apr. 1, 2022. [cited by applicant]
Office Action from corresponding Canadian Patent Application No. 3,033,736, dated Aug. 26, 2022. [cited by applicant]
PCT Application No. PCT/KR2017/008835_International Search Report Written Opinion with its translation, Dec. 20, 2017. [cited by applicant]
PCT Application No. PCT/KR2018/005284_International Search Report with its translation, Aug. 24, 2018. [cited by applicant]
PCT Application No. PCT/KR2018/005284_International Search Report Written Opinion with its translation, Aug. 24, 2018. [cited by applicant]
JP Patent Application No. 2019-561310_Office Action with its translation, Jan. 26, 2021. [cited by applicant]
Australian Patent Application No. 2017308473_Office Action, Jan. 20, 2021. [cited by applicant]
Australian Patent Application No. 2018264636_Office Action, Feb. 25, 2021. [cited by applicant]
RU Patent Application No. 2019106669_Office Action with its translation, Feb. 18, 2021. [cited by applicant]
RU Patent Application No. 2019106669_Search Report with its translation, Feb. 18, 2021. [cited by applicant]
Kuklina E.M., Molecular Mechanisms of T Cell Anergy, Biokhimiya (Biochemistry), vol. 78, 144-156, 2013. [cited by applicant]
Su, S., et al.; “CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients”, Scientific Reports, 2016,6: 20070, pp. 1-14. [cited by applicant]
The Journal of Immunology, vol. 196, Issue 1, Supplement, Immunology. [cited by applicant]
NCBI, GenBank: EF064716.1, Nov. 13, 2006. [cited by applicant]
KR Patent Application No. 10-2017-0103009_Office Action with its translation, Apr. 9, 2018. [cited by applicant]
KR Patent Application No. 10-2017-0103009_Final rejection with its translation, Aug. 2, 2018. [cited by applicant]
KR Patent Application No. 10-2017-0103009_Office Action after Re-examination with its translation, Oct. 8, 2018. [cited by applicant]
KR Patent Application No. 10-2017-0103009_Final rejection after Re-examination with its translation, Apr. 11, 2019. [cited by applicant]
Korean Patent Application No. 10-2019-0068999_Final rejection with its translation, Mar. 10, 2021. [cited by applicant]
Korean Patent Application No. 10-2019-7036278_Office Action with its translation, Apr. 23, 2021. [cited by applicant]
Fagerlund et al. (2015) “The Cpf1 CRISPR-Cas protein expands genome-editing tools.”, [cited by applicant]
International Search Report (ISR) dated Aug. 24, 2018 issued in International Patent Application No. PCT/KR2018/005284, with English Translation. [cited by applicant]
International Search Report (ISR) dated Dec. 20, 2017 issued in International Patent Application No. PCT/KR2017/008835, with English Translation. [cited by applicant]
Office Action from corresponding Japanese Patent Application No. 2019-561310, issued Dec. 17, 2021. [cited by applicant]
Park, J., et al.; “Cas-Designer: a web-based tool for choice of CRISPR-Cas9 target sites”, Bioinformatics, 31(24), 2015, 4014-4016. [cited by applicant]
Notice of Allowance from corresponding Korean Patent Application No. 10-2019-0068999, issued Apr. 21, 2021. [cited by applicant]
Pennisi, E., “The CRISPR Craze”, [cited by applicant]
Chinese Office Action for Application No. 201880045774.2, dated Nov. 24, 2022. [cited by applicant]
Office Action from corresponding Chinese Patent Application No. 201780063250.1, dated Mar. 31, 2023. [cited by applicant]
Office Action from corresponding U.S. Appl. No. 16/324,955, dated May 25, 2023. [cited by applicant]
Office Action from corresponding U.S. Appl. No. 16/324,955, dated Nov. 28, 2023. [cited by applicant]
Mout et al. In vivo delivery of CRISPR/Cas9 for therapeutic gene editing: Progress and Challenges. Bioconjugate Chem. 28:880-884, (Year: 2017). [cited by applicant]
Notice of Allowance from corresponding Chinese Application No. 201780063250.1, dated Jun. 17, 2024. [cited by applicant]