RNA engineered T cells for the treatment of cancer

• US 10,421,960 B2
• Filed: 09/17/2012
• Issued: 09/24/2019
• Est. Priority Date: 09/16/2011
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: June, Carl H., Zhao, Yangbing
• Primary Examiner(s): Bertoglio, Valarie E
• Application Number:
  US14/342,904
• Publication Number:
  US 20140227237A1
• Time in Patent Office: 2,563 Days
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C07K 2319/72 containing SH2 domain
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus

Abstract

The present invention relates to compositions and methods for generating RNA Chimeric Antigen Receptor (CAR) transfected T cells. The RNA-engineered T cells can be used in adoptive therapy to treat cancer.

Claims

1. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector comprises the nucleic acid sequence of SEQ ID NO:

4.

2. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO:

6 and SEQ ID NO;

8.

3. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector comprises the nucleic acid sequence of SEQ ID NO:

5.

4. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO:

7, SEQ ID NO;

9, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23 and SEQ ID NO;

24.

5. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector comprises the nucleic acid sequence of SEQ ID NO:

28.

6. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA, wherein the RNA comprises a nucleic acid sequence encoding an extracellular domain, a transmembrane domain, a costimulatory signaling region, and a signaling domain of CD3-zeta, and wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO:

10, SEQ ID NO;

11, SEQ ID NO;

12 and SEQ ID NO;

13.

7. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA encoding a chimeric antigen receptor (CAR), wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector comprises the nucleic acid sequence of SEQ ID NO:

27.

8. An activated T cell comprising a transfected in vitro transcribed RNA or synthetic RNA, wherein the RNA comprises a nucleic acid sequence encoding an extracellular domain, a transmembrane domain, a costimulatory signaling region, and a signaling domain of CD3-zeta, and wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO:

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17 and SEQ ID NO;

18.