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Technologies

A Method to Inhibit Cancer Metastasis and Eradicate Tumors

ID: 2021-010 Using the power of Junctional Adhesion Molecules combined with the target of CAR-T to inhibit cancer metastasis and eradicate tumors.

Principal Investigators: Christopher Mendoza and Dario Mizrachi

Cancer treatment traditionally involves chemotherapy and surgery. Other promising treatments such as genetically engineered T cells called CAR-T (chimeric antigen receptor T cells) target surface proteins such as CD19. Unfortunately, these treatments do not inhibit metastasis. This invention consists of three components:

  1. A fusion protein that detects cancer cells based on their lower pH with the help of pHLIP.
  2. JAM binding to other tight junction components with neighboring cells that inhibits the metastasis of cancer cells.
  3. A signaling target protein domain for CD19 that allows for the recognition of anit-CD19 CAR-T cells to target and eradicate the tumor.

This invention is the first biologic containing these three separate domains with distinct functions targeting cancer.

The production of a biologic that inhibits metastasis of cancer cells would improve upon currently available targeted cancer treatment. For example, using a fusion protein that would recognize the targeted cell and insert itself into the membrane based on the decreased pH of tumors, which would also allow for the increased interaction between these cancer cells by increasing the tight junctions would decrease the incidence of metastasis. Also, using a targeted signal located on the surface of the membrane of cancer cells that is recognized by existing CAR-T technology would allow for the recognition of a great variety of tumors. Current immune therapy methods are limited due to the need to create individual types of CAR-T cells that recognize different targets such as CD19, CD38 and others, and due to the cost of production. The reason for failure of some of the CAR-T treatments is based on the poor health of the individual, which can result in a low quantity of responsive amount of transformed immune cells.

In our studies, we have purified the four members of the Junctional Adhesion Molecule (JAM) protein family JAM-A, -B, -C and 4 [1]. We have determined that JAMs increase the cell-cell interactions when they are expressed in the tight junction of the cell. We also determined that there is an increase in binding between heterotypic interactions of JAMs compared to homotypic interactions and E-Cadherin [1]. Therefore using the JAM proteins in this technology will allow for the formation of tight junctions in homotypic or heterotypic interactions which will subsequently result in decreasing metastasis in cancer cells. Depending on the target, tissue and the type of cancer that the patient has we will need to use a member of the JAM family such as JAM-A, -B, C or 4, to increase tight junction formation with other members of the JAM family.

Studies have shown that a peptide sequence known as pH-low-insertion peptide (pHLIP) can recognize changes in pH and insert itself into the membrane of cancer cells. The pHLIP inserts its C-terminus through a membrane under low pH conditions (6-6.5) [2,3]. This peptide has been used to deliver compounds such as phalloidin which is linked to the C-terminus and is cleaved inside the cells. This results in the immobilization of cytoskeleton and multinucleation due to F actin binding and filament stabilization [2]. Using this peptide will allow the fusion biologic to target the cancer cell's low-pH environment, and insert itself into the membrane.

Based on the previous studies, we are proposing a method in which we produce a biologic composed of a fusion protein that contains three domains with specific functions. The first domain will consist of a pH-sensitive peptide, pHLIP that will recognize and insert itself into the membrane of cancer cells [2,3]. The second domain will consist of the Junctional Adhesion Molecules (-A, -B, -C and 4) which will increase the number of tight junctions, allowing for cell-cell interactions of the tumor cells that will decrease metastasis. The third domain will consist of the CD19 extracellular region of the target protein that has been seen in many cancers [4,5,6]. This CD19 region will enable the CAR-T cells to recognize, bind to, and eradicate the tumor.

To achieve the goal of decreasing metastasis we have created a biologic consisting of a fusion protein N-CD19-GS Linker-JAM-pHLIP peptide-C. This technology is based on the combination of tight junction components that have been shown in our laboratory to bind tighter than other tight junction components such as claudin proteins. This fusion protein allows the pHLIP peptide to recognize the lower-pH environment produced by cancer cells. When the peptide recognizes the decrease in pH, it will incorporate itself into the membrane of the cancer cell. The JAM region of the fusion protein will bind to neighboring JAMs, promoting tight junction formation, which will result in the inhibition of metastasis of the cancer cells. This approach harnesses the power of tight junctions that allow for the cancer cells to stay within the tumor and not spread throughout the body. The extracellular CD19 region of the fusion protein will be targeted by CAR-T cells, which will eradicate the tumor. The novelty of this approach is the introduction of a target site for CAR-T cells on the biologic that can recognize tumors that do not have CD19 in their membrane surfaces.

The power of cell adhesion can be harnessed to decrease the incidence of metastasis in tumor cells. Using the tight junction protein junctional adhesion molecules, or JAMs (-A, -B, -C and 4), the incidence of metastasis can be decreased. With the combination of pH-sensitive peptides, pHLIP with JAMs, we can decrease metastasis. Additionally, using CD19 in a fusion with JAMs (-A, -B, -C, and 4) will attract CAR-T cells to target these tumors.

Using the power of cell adhesion, this invention will allow for the increase of the tight junction's strength in tumors to prevent metastasis. With the combination of the pHLIP technology, specific tumor cells that will increase cell-cell interactions through tight junctions can be targeted, resulting in a decrease in metastasis of tumor cells. With the help of CD 19 the binding of CAR-T cells can be increased which will target these tumors and eradicate them.

References:

  1. Mendoza C, Nagidi SH, Mizrachi D. Molecular Characterization of the Extracellular Domain of Human Junctional Adhesion Proteins. International Journal of Molecular Sciences. 2021; 22(7):3482. https://doi.orn/10.3390/ijms22073482
  2. An M., Wijesinghe D., Andreev 0. A., Reshetnyak Y. K., Engelman D. M. (2010). pH-(low)insertion-peptide (pHLIP) translocation of membrane impermeable phalloidin toxin inhibits cancer cell proliferation. Proceedings of the National Academy of Sciences Nov 2010, 107 (47) 20246-20250; DOI: 10.1073/pnas.1014403107
  3. Tapmeier, T.T.; Moshnikova, A.; Beech, J.; Allen, D.; Kinchesh, P.; Smart, S.; Harris, A.; McIntyre, A.; Engelman, D.M.; Andreev, O.A.; et al. The pH low insertion peptide pHLIP Variant 3 as a novel marker of acidic malignant lesions. Proc. Natl. Acad. Sci. USA 2015, 112, 9710-9715.
  4. Maude, S. L., Teachey, D. T., Porter, D. L., & Grupp, S. A. (2015). CD19-targeted chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Blood, 125(26), 4017-4023. httos://doi.or!!/1 0. l 182/blood-2014-12-580068
  5. Drent, E., Groen, R. W., Noort, W. A., Themeli, M., Lammerts van Bueren, J. J., Parren, P. W., Kuball, J., Sebestyen, Z., Yuan, H., de Bruijn, J., van de Donk, N:w., Martens, A. C., Lokhorst, H. M., & Mutis, T. (2016). Pre-clinical evaluation of CD38 chimeric antigen receptor engineered T cells for the treatment of multiple myeloma. Haematologica, 101 (5), 616-625. https://doi.oru/10.3324/haematol.2015 .137620
  6. Guo, Y., Feng, K., Tong, C. et al. Efficiency and side effects of anti-CD38 CART cells in an adult patient with relapsed B-ALL after failure of bi-specific CD19/CD22 CART cell treatment. Cell Mol Immunol 17, 430-432 (2020). https://doi.org/10.1038/s41423-019-0355-5

For more information, contact Mike Alder (801-422-3049)

Links and Resources

  1. One Page Summary PDF
  2. Inventor Webpage - Dario Mizrachi
  3. Mizrachi Lab Webpage