Epidermal Derived Neural Crest Stem Cells for Treatment of Demyelinating Diseases.

(NIBIB, R01 EB023114 at NCE; R01 NS130130-01A1 under review)

Stelios T. Andreadis Lab

Introduction & Significance

The Andreadis laboratory discovered a population of neural crest like stem cells in cultures of postnatal human epidermal keratinocytes (they were termed as KC-NC) by mimicking signaling events that occur at the neural plate border.

During development, neural crest cells (NC) are induced by signaling events at the neural plate border of all vertebrate embryos.  Initially arising within the central nervous system, NC cells subsequently undergo an epithelial to mesenchymal transition to migrate into the periphery, where they differentiate into diverse cell types.

Research Findings

Zoom image: Fig. 1: Keratinocyte-derived neural crest like stem cells (KC-NC) stem cells and their derivatives including neurons, Schwann cells and melanocytes.  3.1.

Fig. 1: Keratinocyte-derived neural crest like stem cells (KC-NC) stem cells and their derivatives including neurons, Schwann cells and melanocytes. 

The Andreadis laboratory discovered a population neural crest like stem cells in cultures of postnatal human epidermal keratinocytes (they were termed as KC-NC) by mimicking signaling events that occur at the neural plate border (Fig. 1). Genomewide transcriptome analyses showed that keratinocyte-derived NC cells were similar to those derived from human embryonic stem cells.  Moreover, they gave rise in vitro and in vivo to neural crest derivatives such as peripheral neurons, melanocytes, Schwann cells and mesenchymal cells (osteocytes, chondrocytes, adipocytes and smooth muscle). Upon transplantation into chicken embryos, KC-NC migrated along stereotypical pathways and gave rise to multiple NC derivatives 1. 

In subsequent studies, the lab identified the factors that promote expansion of KC-NC and maintain KC-NC multipotency 2-4. They also demonstrated that the same signaling factors can be used to obtain multipotent and functional KC-NC from the epidermis of elderly donors. Interestingly, KC-NC from older donors exhibited significantly younger epigenetic age than epidermal KC, perhaps indicating greater potential for cell therapies 5.

Given the accessibility, high proliferative capacity and ease of reprogramming, KC-NC represent a potentially useful source of functional therapeutic cells for regenerative medicine and tissue engineering applications. They can also provide an excellent culture system for studying human disease, similar to induced pluripotent stem cells but without the need for genetic modification or reprogramming to the pluripotent state.

Zoom image: Fig. 2(A) : Injectable shear thinning hydrogel (STH) enhanced cell survival and cell number of human oligodendrocyte progenitors transplanted into the corpus callosum of mouse model of demyelinating disease.  3.2A.

Fig. 2(A) : Injectable shear thinning hydrogel (STH) enhanced cell survival and cell number of human oligodendrocyte progenitors transplanted into the corpus callosum of mouse model of demyelinating disease. 

Zoom image: Fig 2(B): Immunostaining for Myelin Basic Protein (MBP; green) and hNA (red) to mark human cells; DAPI stained nuclei (blue).  3.2B.

Fig 2(B): Immunostaining for Myelin Basic Protein (MBP; green) and hNA (red) to mark human cells; DAPI stained nuclei (blue). 

Future Work

Current and future work in the Andreadis laboratory aims at utilizing these stem cells for treatment of neuro-degenerative and demyelinating disorders. To this end, we designed novel shear thinning hydrogels to enhance survival and function of transplanted KC-NC and their Schwann cell derivatives to the brain of the Shiverer mouse, an established model of demyelinating diseases (Fig. 2).  

Students of the Project

Post-doc: Mohamed Alaa Mohamed, Ph.D.

Graduate students: Pihu Mehrotra; Ashis Kumar Podder

Collaborators: Fraser Sim (UB Pharmacology); Marianne Bronner (Caltech)

References

  1. Bajpai, V.K., Kerosuo, L., Tseropoulos, G., Cummings, K.A., Wang, X., Lei, P., Liu, B., Liu, S., Popescu, G.K., Bronner, M.E. & Andreadis, S.T. Reprogramming Postnatal Human Epidermal Keratinocytes Toward Functional Neural Crest Fates. Stem cells 35, 1402-1415 (2017).
  2. Mehrotra, P., Ikhapoh, I., Lei, P., Tseropoulos, G., Zhang, Y., Wang, J., Liu, S., Bronner, M.E. & Andreadis, S.T. Wnt/BMP mediated metabolic and epigenetic reprogramming preserves multipotency of skin derived neural crest like stem cells. Stem Cells In revision(2022).
  3. Mehrotra, P., Tseropoulos, G., Bronner, M.E. & Andreadis, S.T. Adult tissue-derived neural crest-like stem cells: Sources, regulatory networks, and translational potential. Stem cells translational medicine 9, 328-341 (2020).
  4. Tseropoulos, G., Moghadasi Boroujeni, S., Bajpai, V.K., Lei, P. & Andreadis, S.T. Derivation of neural crest stem cells from human epidermal keratinocytes requires FGF-2, IGF-1, and inhibition of TGF-beta1. Bioeng Transl Med 3, 256-264 (2018).
  5.  Moghadasi Boroujeni, S., Koontz, A., Tseropoulos, G., Kerosuo, L., Mehrotra, P., Bajpai, V.K., Selvam, S.R., Lei, P., Bronner, M.E. & Andreadis, S.T. Neural crest stem cells from human epidermis of aged donors maintain their multipotency in vitro and in vivo. Sci Rep 9, 9750 (2019).