OUR SCIENCE:

Notch Signaling and T Cell Differentiation.

Notch Therapeutics’ unique technologies and capabilities are rooted in foundational discoveries in developmental biology.

Notch cell therapies are based on more than a century of milestones in stem cell and Notch signaling research.

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It has been more than 100 years since scientists first described unusual fruit flies born with “notched” wings that looked as though a small bite had been taken out of them. When the causal gene of this trait was cloned in the 1980s, it was named “Notch” and hypothesized to code for a cell membrane protein involved in cell-cell interactions.

We know today that the Notch genes encode a family of receptors that drive cell fate decisions by binding ligands and transmitting signals between adjacent cells. The Notch Signaling Pathway is a conserved and critical component of development for most organs and tissues, and mutations in related genes result in a wide range of phenotypes ranging from “notched” wings in fruit flies to various congenital disorders in humans.

In the late 1990s, it was discovered that bone marrow cells from mice carrying mutations in the Notch1 gene could give rise to all hematopoietic lineages except T cells. Shortly thereafter, Notch Therapeutics scientific co-founder, Juan Carlos Zúñiga-Pflücker, made the seminal discovery that by artificially expressing Notch ligands (Delta-like, DL) in mouse OP9 stromal cells and co-culturing those OP9-DL cells with hematopoietic progenitor cells, one could finally generate human T cells in a dish without a thymus. Dr. Zúñiga-Pflücker has continued to advance strategies to generate T cells from stem cells.

Although the OP9-DL cell system has been widely adopted for research purposes, its animal origins and lack of scalability have prevented its use in therapeutics. Furthermore, the mechanisms of action of the feeder cell line are unknown, since providing Notch ligands alone to developing T cells fails to robustly induce T cell differentiation. The novel discovery by Notch co-founders Shreya Shukla, Zúñiga-Pflücker, and Peter Zandstra that the cell-surface molecule, VCAM-1, enhances Notch signaling to enable the scalable production of T cells from any source of hematopoietic progenitor cells led to the development of Notch’s proprietary ”Engineered Thymic Niche” technology.

Induced Pluripotent Stem Cells.

Current T cell therapies require a complex, patient-specific manufacturing process. This “vein-to-vein” processing of a CAR-T therapy leads to delays and variability that negatively impact patient outcomes. To fulfill the promise of a cell therapy that can be delivered as an off-the-shelf, fully characterized medicine, Notch Therapeutics is manufacturing T cells from induced pluripotent stem cells (iPSCs).

iPSCs were discovered in 2006 by Shinya Yamanaka, who was later awarded the Nobel Prize for the discovery that terminally differentiated mature cells can be reprogrammed to once again have the capability to give rise to any cell of the body when given the appropriate developmental signals. Since iPSCs can be cultured at massive scale in their undifferentiated state, they serve as a potentially limitless source of starting materials for the production of therapeutically relevant cells.

Synthetic Biology.

The fact that iPSCs can be cultured at massive scale makes it possible to modify the genome of the cells to encode desirable attributes that will be present and active in the final differentiated cell product. For example, iPSCs can be modified to control stem cell division and differentiation, alter the tumor microenvironment, and prevent immune activity or susceptibility of the final product.