PI: Dr. Vincenzo Cirulli
Location: Institute for Stem Cell & Regenerative Medicine (ISCRM)
Website: http://depts.washington.edu/iscrm/research/faculty/vincenzo-cirulli

Netrin protein nanomaterials as regulators of stem cell differentiation toward pancreatic β-cells

Cell interactions with the extracellular microenvironment play critical roles in tissue morphogenesis and homeostasis. In the pancreas, development of insulin-producing islet cells (b-cells) depends on a series of highly regulated processes that include branching morphogenesis, proliferation, migration and differentiation of progenitors into the surrounding mesenchyme where they organize into cell clusters (islets of Langerhans). Despite significant advances in our understanding of pathways regulating the specification, expansion and differentiation of the islet cell lineage, mechanisms governing the recruitment and delamination of pancreatic progenitors from the ductal tree remain largely unknown. Among components of the extracellular microenvironment that can significantly affect cellular development and function, secreted factors such as Netrins are of special interest since they regulate cell adhesion and migration through mechanisms of “chemo-attraction” and “chemo-repulsion”. Our laboratory has demonstrated that Netrins mediate the adhesion and migration of PDX-1+pancreatic progenitors through recruitment of integrin receptors, and promote insulin and glucagon gene expression. These studies provided the first evidence that neural chemo-tropic factors are important regulators of pancreatic progenitors navigation and differentiation into insulin-producing b-cells.

Recently, we have extended our analysis of Netrins’ function(s) to stem cells with the ultimate goal of uncovering possible instructive properties of these neural chemotropic factors in stem cell fate determination. In preliminary experiments we observed that Netrins are able to impart unique developmental decisions at distinct stages of stem cell differentiation toward pancreatic cell lineages.

In light of these observations, building on the established expertise of the Institute for Protein Design at The University of Washington, we aim to bioengineer synthetic protein sheets that recapitulate in vivo-like extracellular niches containing combination of select ECMs and Netrins’ domains, and that can be used to foster the differentiating of stem cells toward the pancreatic islet cell lineage.

Collectively, we anticipate that this line of research may lead to the design of innovative strategies for the derivation of insulin-producing cells from stem cell preparations, and ultimately contribute to the development of novel cell-based replacement therapies to cure diabetes.

Relevant Literature from the sponsor laboratories

Cirulli V., Beattie, G.M., Klier, G., Ellisman, M., Ricordi, C., Quaranta, V., Frasier, F., Ishii, J.K., Hayek, A., Salomon, D.R. (2000). Expression and function of alpha(v)beta(3) and alpha(v)beta(5) integrins in the developing pancreas: roles in the adhesion and migration of putative endocrine progenitor cells. J. Cell Biol. 150, 1445-1460.

Cirulli V, Crisa, L, Beattie, GM, Mally, MI, Lopez, AD, Fannon, A, Ptasznik, A, Inverardi, L, Ricordi, C., Deerinck, T. Ellisman, M., Reisfeld, R.A., and A. Hayek. The KSA antigen (EpCAM) mediates cell-cell adhesion of pancreatic epithelial cells: morphoregulatory roles in pancreatic islet development. J Cell Biol 140: 140:1519-1534, 1998.

Yebra M., Montgomery A.M.P., Diaferia G.R., T. Kaido, Silleti S., Perez B., Just M.L., Hildbrand S., Hurford R., Florkiewcz E., Tessier-Lavigne M., and Cirulli V. “Integrin-mediated recognition of the neural chemo-attractant Netrin-1: a novel adhesive and migratory cue for epithelial cells.” Developmental Cell, 5:695-707, 2003.

Yebra M, Diaferia GR, Montgomery AM, Kaido T, Brunken WJ, Koch M, Hardiman G, Crisa L, Cirulli V. Endothelium-derived Netrin-4 supports pancreatic epithelial cell adhesion and differentiation through Integrins α2β1 and α3β1. PLoS One. 2011;6(7):e22750. Epub 2011 Jul 29.

King NP, Bale JB, Sheffler W, McNamara DE, Gonen S, Gonen T, Yeates TO, Baker D. Accurate design of co-assembling multi-component protein nanomaterials. Nature. 2014 Jun 5;510(7503):103-8. doi: 10.1038/nature13404. Epub 2014 May 25.