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US-12624102-B2 - CD147 chimeric antigen receptors and methods of use

US12624102B2US 12624102 B2US12624102 B2US 12624102B2US-12624102-B2

Abstract

Modified single chain variable fragments (scFv) that specifically bind CD147 are provided. Also provided are chimeric antigen receptors (CARs) including the modified CD147 scFv, nucleic acids encoding the CARs, vectors including the nucleic acids encoding the CARs, and immune cells expressing the CARs. Methods of treating a subject with cancer including administering to the subject an immune cell expressing a disclosed CD147-CAR are also provided.

Inventors

  • Dongfang Liu

Assignees

  • RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY

Dates

Publication Date
20260512
Application Date
20200228

Claims (20)

  1. 1 . A nucleic acid molecule encoding a chimeric antigen receptor comprising: (a) an antigen binding domain that specifically binds CD147, wherein the antigen binding domain is encoded by a nucleic acid comprising at least 95% sequence identity to SEQ ID NO: 1, and comprises an amino acid sequence comprising the variable heavy chain (VH) domain complementarity determining region 1 (CDR1), CDR2 and CDR3 amino acid sequences of amino acid positions 26-32, 52-59, and 101-104 of SEQ ID NO: 2, respectively, and the variable light chain (VL) domain CDR1, CDR2 and CDR3 amino acid sequences of amino acid positions 155-165, 181-187, and 220-228 of SEQ ID NO: 2, respectively; (b) a hinge domain; (c) a transmembrane domain; and (d) an intracellular domain comprising one or more co-stimulatory molecule intracellular domains and an intracellular signaling domain.
  2. 2 . The nucleic acid molecule of claim 1 , wherein the antigen binding domain has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2 or comprises the amino acid sequence of SEQ ID NO: 2.
  3. 3 . The nucleic acid molecule of claim 1 , wherein the one or more co-stimulatory molecule intracellular domains comprise intracellular domains of CD28 and 4-1BB, and/or the intracellular signaling domain comprises a signaling domain of CD3ζ, and/or the hinge domain comprises an IgG1 hinge domain, and/or the transmembrane domain comprises a CD28 transmembrane domain or a CD8a transmembrane domain.
  4. 4 . The nucleic acid molecule of claim 1 , wherein the chimeric antigen receptor comprises an amino acid sequence with at least 98% identity to the amino acid sequence of SEQ ID NO: 5 or comprises the amino acid sequence of SEQ ID NO: 5.
  5. 5 . The nucleic acid molecule of claim 1 , wherein the chimeric antigen receptor further comprises one or more additional antigen binding domains, and/or an inducible suicide molecule, and/or a cytokine receptor intracellular domain.
  6. 6 . The nucleic acid molecule of claim 5 , wherein the chimeric antigen receptor comprises an amino acid sequence with at least 98% identity to the amino acid sequence of SEQ ID NO: 7 or comprises the amino acid sequence of SEQ ID NO: 7.
  7. 7 . The nucleic acid molecule of claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 14.
  8. 8 . The nucleic acid molecule of claim 1 , wherein the antigen binding domain is encoded by a nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 1.
  9. 9 . A nucleic acid molecule, comprising at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 1, and encoding an antigen binding domain that specifically binds CD147, wherein the antigen binding domain comprises an amino acid sequence comprising the variable heavy chain (VH) domain complementarity determining region 1 (CDR1), CDR2 and CDR3 amino acid sequences of amino acid positions 26-32, 52-59, and 101-104 of SEQ ID NO: 2, respectively, and the variable light chain (VL) domain CDR1, CDR2 and CDR3 amino acid sequences of amino acid positions 155-165, 181-187, and 220-228 of SEQ ID NO: 2, respectively.
  10. 10 . A vector comprising the nucleic acid molecule of claim 1 .
  11. 11 . The vector of claim 10 , further comprising an inducible promoter or enhancer nucleic acid molecule operably linked to the nucleic acid molecule.
  12. 12 . The vector of claim 11 , wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 15.
  13. 13 . A T cell, natural killer (NK) cell, natural killer T (NKT) cell, double negative T (DNT) cell, neutrophil, or macrophage comprising the nucleic acid molecule of claim 1 .
  14. 14 . A T cell, NK cell, natural killer T (NKT) cell, double negative T (DNT) cell, neutrophil, or macrophage comprising the vector of claim 10 .
  15. 15 . An immune cell, comprising the nucleic acid molecule of claim 1 .
  16. 16 . The nucleic acid molecule of claim 9 , comprising the nucleic acid sequence of SEQ ID NO: 1.
  17. 17 . An immune cell, comprising a chimeric antigen receptor comprising: (a) an antigen binding domain that specifically binds CD147, wherein the antigen binding domain is encoded by a nucleic acid comprising at least 95% sequence identity to SEQ ID NO: 1, and comprises an amino acid sequence comprising the variable heavy chain (VH) domain complementarity determining region 1 (CDR1), CDR2 and CDR3 amino acid sequences of amino acid positions 26-32, 52-59, and 101-104 of SEQ ID NO: 2, respectively, and the variable light chain (VL) domain CDR1, CDR2 and CDR3 amino acid sequences of amino acid positions 155-165, 181-187, and 220-228 of SEQ ID NO: 2, respectively; (b) a hinge domain; (c) a transmembrane domain; and (d) an intracellular domain comprising one or more co-stimulatory molecule intracellular domains and an intracellular signaling domain.
  18. 18 . The immune cell of claim 17 , wherein the antigen binding domain is encoded by a nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 1.
  19. 19 . The immune cell of claim 18 , wherein the cell is a T cell, natural killer (NK) cell, natural killer T (NKT) cell, double negative T (DNT) cell, neutrophil, or macrophage.
  20. 20 . A method of producing CD147-CAR-T cells or CD147-CAR-NK cells, comprising transducing or transfecting T cells or NK cells with the vector of claim 10 .

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is the § 371 U.S. National Stage of International Application No. PCT/US2020/020436, filed Feb. 28, 2020, which was published in English under PCT Article 21(2), which in turn claims the benefit of U.S. Provisional Patent Application No. 62/819,403, filed Mar. 15, 2019, which is incorporated herein by reference in its entirety. ACKNOWLEDGMENT OF GOVERNMENT SUPPORT This invention was made with government support under grant numbers HL150852 and AI130197 awarded by the National Institutes of Health. The government has certain rights in the invention. FIELD This disclosure related to immunotherapies, particularly chimeric antigen receptors targeting CD147 and their use for treating cancer. SEQUENCE LISTING INCORPORATION BY REFERENCE The Sequence Listing is submitted as an ASCII text file in the form of the file named Sequence_Listing-2.txt, which was created on Dec. 15, 2025, and is 61,738 bytes, which is incorporated by reference herein. BACKGROUND Liver cancer is the second most common cause of cancer-related death worldwide. The burden of liver cancer is projected to be over 1 million cases by 2030. Liver cancer ranks fifth in terms of global cases and second in terms of deaths for males. More than half a million patients die from hepatocellular carcinoma (HCC) each year. Primary liver cancer includes hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (iCCA), fibrolamellar carcinoma, and hepatoblastoma. HCC and iCCA are the most common primary liver cancers, which account for more than 99% of primary liver cancer cases. HCC alone (nearly 800,000 new cases per year) accounts for 90% of all cases of primary liver cancer. Currently, there is no effective therapy available to treat HCC. Sorafenib (CheckMate-040, a multi-kinase inhibitor widely used for advanced HCC patients with low efficacy and severe side effects) is a first-line standard systemic agent for HCC. Currently, PD-1 blockade Opdivo (Nivolumab) has been approved by the US Food and Drug Administration (FDA) as a second line treatment strategy for patients with HCC who have been previously treated with Sorafenib. Clinical trials testing PD-1 blockade as a first-line treatment for HCC are underway. Meanwhile, various clinical trials using PD-1 or PD-L1 blockades in combination with other interventions are ongoing as well. For example, a study evaluating anti-PD-1 antibody in combination with anti-CTLA-4 antibody in patients with resectable and potentially resectable HCC is being tested in clinical trials (NCT03222076). Chimeric antigen receptor (CAR)-modified T cell therapy has become a promising immunotherapeutic strategy for the treatment of various blood cancers. Despite recent advances in CAR-modified T cell immunotherapy in blood cancers, high costs and severe toxicity have hindered its widespread use. Meanwhile, CAR-T cells face additional challenges during the targeting of solid tumors, such as maintaining durable proliferation and persistence in the tumor microenvironment. An additional challenge for CAR-mediated immunotherapy for liver cancer is to find an effective target. SUMMARY CD147 is expressed on different cell types (e.g., hematopoietic, epithelial, and endothelial cells) at varying levels. However, CD147 is significantly upregulated in disease states, such as in HCC, breast cancer, bladder cancer, colorectal cancer, ovarian cancer, melanoma, and osteosarcoma. CARs that specifically target cells expressing CD147 are provided. These CARs can be used in immunotherapy of cancers expressing or overexpressing CD147. Disclosed herein are modified single-chain variable fragments (scFvs) that specifically bind CD147. In some embodiments, the scFv has an amino acid sequence that includes the variable heavy chain (VH) domain complementarity determining region 1 (CDR1), CDR2 and CDR3 amino acid sequences of SEQ ID NO: 8 and the variable light chain (VL) domain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NO: 9. In some examples, the scFv has at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 2 or includes or consists of the amino acid sequence of SEQ ID NO: 2. Also provided are nucleic acids that encode the modified CD147 scFv, such as a nucleic acid with at least 90% sequence identity to the nucleic acid molecule of SEQ ID NO: 1, or include or consist of the nucleic acid sequence of SEQ ID NO: 1 and vectors including the nucleic acid sequence. In additional embodiments, provided are vectors encoding the modified CD147 scFv (such as SEQ ID NO: 1), which further comprise an inducible promoter or enhancer nucleic acid molecule operably linked to the CD147 scFv nucleic acid molecule. In some examples, the enhancer nucleic acid is a Gal4 upstream activation sequence (UAS) that is operably linked to a nucleic acid encoding the CD147 scFv. In another example, the vector is a synNotch construct, for example a vector including the nucleic acid sequence of th