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US-12624085-B2 - Compositions and methods of NKG2D chimeric antigen receptor T cells for controlling triple-negative breast cancer

US12624085B2US 12624085 B2US12624085 B2US 12624085B2US-12624085-B2

Abstract

The invention includes a chimeric antigen receptor (CAR) specific for a natural-killer group 2, member D ligand (NKG2DL), compositions comprising the CAR, polynucleotides encoding the CAR, vectors comprising a polynucleotide encoding the CAR, and recombinant T cells comprising the CAR. The invention also includes methods of making a genetically modified T cell expressing a NKG2D-CAR wherein the expressed CAR comprises a NKG2D extracellular domain. The present invention also relates generally to the use of T cells engineered to express a CAR to treat cancer associated with NKG2D.

Inventors

  • Daniel Powell
  • De-Gang SONG

Assignees

  • THE TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA

Dates

Publication Date
20260512
Application Date
20190624

Claims (15)

  1. 1 . A nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain, wherein the CAR comprises, from amino to carboxy terminus, the NKG2D extracellular domain, the CD27 costimulatory domain, and the CD3 zeta signaling domain, and wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15.
  2. 2 . The nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 14.
  3. 3 . A vector comprising the nucleic acid molecule of claim 1 .
  4. 4 . A chimeric antigen receptor (CAR) comprising a natural-killer group 2, member D (NKG2D) extracellular domain, a CD27 costimulatory domain, and a CD3 zeta signaling domain, wherein the CAR comprises, from amino to carboxy terminus, the NKG2D extracellular domain, the CD27 costimulatory domain, and the CD3 zeta signaling domain, and wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15.
  5. 5 . The CAR of claim 4 , wherein the CAR is encoded by the nucleic acid sequence of SEQ ID NO: 14.
  6. 6 . A genetically modified T cell comprising the CAR of claim 4 .
  7. 7 . A method for treating cancer in a subject, the method comprising: administering to the subject an effective amount of the genetically modified T cell of claim 6 , thereby treating cancer in the subject.
  8. 8 . A genetically modified T cell comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding, from amino to carboxy terminus, a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain, wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15 or is encoded by the nucleic acid sequence of SEQ ID NO: 14.
  9. 9 . A genetically modified T cell comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 16 or encodes the amino acid sequence of SEQ ID NO: 17.
  10. 10 . A genetically modified T cell comprising a chimeric antigen receptor (CAR), wherein the CAR comprises the amino acid sequence of SEQ ID NO: 19 or is encoded by the nucleic acid sequence of SEQ ID NO: 18.
  11. 11 . A vector comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding, from amino to carboxy terminus, a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain, wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 1.
  12. 12 . A method for treating triple negative breast cancer (TNBC) in a subject, the method comprising: administering to the subject an effective amount of a genetically modified T cell comprising a nucleic acid molecule encoding, from amino to carboxy terminus, a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence of a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain, thereby treating TNBC in the subject.
  13. 13 . The method of claim 12 , wherein the subject is a human.
  14. 14 . The method of claim 12 , further comprising wherein the subject is administered a secondary treatment.
  15. 15 . The method of claim 14 , wherein the secondary treatment is selected from the group consisting of immune checkpoint blockade, chemotherapy, radiation, and surgery.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS The present application is a 35 U.S.C. § 371 national phase application from, and claims priority to, International Application No. PCT/US2019/038739, filed Jun. 24, 2019, and published under PCT Article 21(2) in English, which claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62/689,552, filed Jun. 25, 2018, all of which applications are incorporated herein by reference in their entireties. BACKGROUND OF THE INVENTION Triple negative breast cancer (TNBC), an aggressive form of breast cancer that lacks significant expression of the human epidermal growth factor receptor 2 (HER2), estrogen receptor (ER) and progesterone receptor (PR), accounts for approximately 15%-20% of invasive breast cancers. In the absence of obvious targets, patients with TNBC do not benefit from endocrine therapy or other available targeted agents. To date, the standard treatment still depends on surgery and adjuvant chemotherapy and radiotherapy. Patients with TNBC have a worse outcome after chemotherapy, compared to breast cancers patients with other subtypes, a finding that reflects the intrinsically adverse prognosis associated with the disease. Thus, effective therapeutic strategies are urgently needed for TNBC patients. Cancer cells, including TNBC cells, frequently upregulate “stress” induced ligands recognized by the NK cell activating receptors NKG2D (natural-killer group 2, member D) and DNAM-1(CD226). The feasibility of targeting NKG2D ligands (NKG2DLs) utilizing a chimeric antigen receptor (CAR) engineered T cell approach was demonstrated by Sentman and colleagues in 2005, and early clinical trial results now show the significant promise of this approach (Sallman et al., 27 Apr. 2018: Haematologica, haematol. 2017.18674). This CAR construct contained the full-length NKG2D fused to the cytoplasmic domain of CD3z with costimulation provided endogenously by Dap10. However, there is no evidence in the art that this CAR is an effective targeted therapy for triple negative breast cancers (TNBC). There is an urgent need in the art for achieving a more specific and effective treatment for triple negative breast cancer (TNBC). The present invention addresses this need. SUMMARY OF THE INVENTION As described herein, the present invention relates to compositions and methods utilizing NKG2D chimeric antigen receptor T cells for controlling cancer (e.g. triple-negative breast cancer). One aspect of the invention includes a nucleic acid molecule encoding a chimeric antigen receptor (CAR). The nucleic acid molecule comprises a nucleic acid sequence encoding a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain. Another aspect of the invention includes a chimeric antigen receptor (CAR) comprising a natural-killer group 2, member D (NKG2D) extracellular domain, a CD27 costimulatory domain, and a CD3 zeta signaling domain. Yet another aspect of the invention includes a genetically modified T cell comprising any one of the CARs disclosed herein. Another aspect of the invention includes a genetically modified T cell comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain. Still another aspect of the invention includes a genetically modified T cell comprising a nucleic acid molecule encoding a CAR, wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 14 or SEQ ID NO: 16 or SEQ ID NO: 18. In another aspect, the invention includes a genetically modified T cell comprising a CAR, wherein the CAR comprises the amino acid sequence of SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19. Still another aspect of the invention includes a vector comprising any one of the nucleic acid molecules disclosed herein. Another aspect of the invention includes a vector comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the nucleic acid molecule comprises a nucleic acid sequence encoding a natural-killer group 2, member D (NKG2D) extracellular domain, a nucleic acid sequence encoding a CD27 costimulatory domain, and a nucleic acid sequence encoding a CD3 zeta signaling domain. Yet another aspect of the invention includes a method for treating cancer in a subject. The method comprises administering to the subject an effective amount of any one of the genetically modified T cells disclosed herein, thereby treating cancer in the subject. Another aspect of the invention includes a method for treating cancer in a subject, the method comprising: administering to the subject an effective amount of a genetically modi