Allogeneic Hematopoietic Stem Cell Transplantation is a curative therapy for many blood disorders, including leukemias and inherited blood diseases. It involves depleting a patient’s diseased bone marrow by cytotoxic therapies and then regenerating a healthy hematopoietic system by transplantation of healthy Hematopoietic Stem Cells (HSCs).

UNMET NEED
Cord blood is an attractive source for stem cell transplants due to easier HLA matching and reduced incidences of GvHD. One of the drawbacks of cord blood is low HSC numbers in publicly accessible cord blood units, which is associated with high graft failure rates.
Current approaches to overcome this drawback is to expand HSCs in culture, as in the case with Gamida Cell’s Omisirge®, where HSCs are expanded with Nicotinamide for 3 weeks in the laboratory and then injected into a patient. This expensive and challenging method of ex vivo expansion puts HSCs in a proliferative state and may exhaust them.

OUR SOLUTION
We employ an alternative strategy where we increase the functionality of HSCs, rather than their numbers. A brief pulse treatment (24 hours) of HSCs with Forskolin/IBMX cocktail that elevates cAMP levels inside HSCs, maintains mitochondrial membrane integrity and upregulates pro-survival MCL1 protein levels thus promoting HSC survival. We also found that, pre-treatment of HSCs with Forskolin/IBMX cocktail enhances the homing of HSCs as seen by increased expression of homing marker CXCR4 at both mRNA and protein level. Finally, we observed enhanced bone marrow repopulating ability of Forskolin/IBMX treated HSCs compared to control treated group in vivo. Brief pulse treatment of HSPCs with Forskolin/ IBMX, showed increased human chimerism in mouse bone marrow even after 24 weeks.

APPLICATIONS
Cord blood transplantation is an effective cure for a wide variety of malignant and non-malignant hematological diseases (mostly in children).
Other potential uses for Forskolin/ IBMX treated HSCs include facilitation of gene editing technologies in HSCs. We anticipate that it will enhance the engraftment and survival of genetically modified HSCs and allow the treatment for the wide spectrum of inherited blood diseases.

STATUS
We’re currently evaluating the transcriptional response to Forskolin/IBMX in HSPCs at single cell level, to identify the molecular regulators that enhance HSC function and find novel marker genes. Future work will also focus to evaluate if Forskolin/IBMX cocktail can enhance the engraftment of lentiviral transduced HSCs.

INTELLECTUAL PROPERTY
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REFERENCES
1) Muddineni, SSNA, Katz Even C, Zipin-Roitman A, Beider K, Raz Y, Weizman E, Nagler A, and Milyavsky M. Mesenchymal stromal cells regulate human hematopoietic stem cell survival and regeneration via cAMP/PKA pathway. BioRxiv 2023 doi.org/10.1101/2023.11.09.566361.