Day 1 :
International University of Health and Welfare, Japan
Keynote: Neuronal differentiation of skin-derived precursors by intracellular delivery of synthesized peptides derived from BC-box proteins
Time : 10:00 - 10:45
Hiroshi Kanno is the Professor of Neurosurgery at International University of Health and Welfare, School of Medicine in Japan. His research interest has been focusing on stem cells and regenerative medicine for 20 years.
Several somatic stem cells have potential to differentiate to neurons, and they are hopeful to be used as grafted donor cells for neuronal regenerative therapy. However, the grated cells little survive and differentiate to functional neurons in recipient neural tissue. To overcome the problem, neural induction using neuro-trophic factors or gene transfer has been employed before grafting, but neuro-trophic factors do not occur specific neural induction, while gene transfer has risk of vector. If a neural induction domain for somatic stem cells exists in proteins to induce neural differentiation, its identification can contribute to neuronal regenerative therapy through neural induction of somatic stem cells using the neural induction domain. We previously demonstrated that von Hippel-Lindau tumor suppressor (VHL) protein has a function of neural induction in skin-derived precursors (SKPs) without any neuro-trophic factors. Then, we hypothesized that a neural induction domain potentially exists in the VHL protein. Here we identify a neural induction domain for somatic stem cells in the VHL protein and show neural induction of the cells by transfer of the domain peptide linked to protein transduction domain (PTD). The neural induction domain in the VHL protein contains BC-box motif [(A,P,S,T)LXXX (A,C) XXX(A,I,L,V)] corresponding to binding site of elongin BC, which is evolutionally conserved from virus to mammalian. Therefore, we proposed that other BC-box proteins also contain the neural induction domain and subsequently show to identify the neural induction domains at amino-acid sequences encoded by BC-box motif within BC-box proteins responsible for neuronal differentiation of somatic stem cells. In addition, we show that the domain has the same function for other somatic stem cells except for neural stem cells. Furthermore, when the domain peptide-transferred stem cells are grafted into recipient nervous tissue in neuronal disease models, the grafted cells differentiate to neurons and neuronal repair for neuronal disease models is achieved. Thus, a neural induction domain is identified at BC-box motif in BC-box proteins. The neuronal differentiation of somatic stem cells is caused by intracellular delivery of the neural induction domain peptide linked to PTD and would contribute to neuronal regenerative therapy.
ReeLabs Pvt. Ltd., India
Keynote: Innovations in regenerative medicine that improve the results of stem cell treatment: 3D cultivation, in vivo modeling of stem cell niche and prenatal stem cells exosomes
Time : 11:00 - 11:45
Abhijit Bopardikar is the Promoter and Founder Director of ReeLabs Pvt. Ltd, Mumbai and Vadivarhe Speciality Chemicals Ltd. (V.S.C.L.), Vadivarhe, Nasik, since 2009.
Statement of the Problem: Hair loss is currently being addressed by hair follicle transplantation. However, for large areas of alopecia, the patient does not have enough hair for auto-transplantation. Proliferation and differentiation of stem cells (SCs) requires a specific microenvironment – “stem cells niche”. For in vivo modulation of organ-specific niches during SCs transplantation could be useful fetal tissue extracts (FTEs). Exosomes are small vesicles that are secreted by various cell types, including SCs. Exosomes can be carried to distant sites via biological fluids and may therefore induce the phenotypic modifications in recipient cells.
Methodology & Theoretical Orientation: Multiplying SCs of hair follicles in 2D culture and introduce them into the scalp skin to form De novo hair follicles were unsuccessful. We developed a technology for creating new hair follicles from SCs in 3D cultures. Also, we investigated the content of growth factors in FTEs and studied the efficacy of FTEs in patients who did not respond to SCs treatment. Finally, we created rejuvenation program, which includes SCs transplantation and exosomes of SCs administration.
Findings: The SCs were transferred to a 3D culture where the formation of primary hair follicles suitable for transplantation occurred under the influence of a specific combination of growth factors. We showed high efficacy of using FTEs for modeling the SCs niche in treatment of liver cirrhosis and non-healing wound in patients who did not have positive response to previous SCs treatment. Transplantation of prenatal hepatoblasts, hematopoietic SCs and fetal liver extracts administration showed efficacy in 75% of liver cirrhosis cases that was characterized by significant decrease of liver fibroscan density, decrease of portal hypertension and ascites, decrease or normalization of biochemical markers of liver damage. In patients with chronic non-healing wounds administration of FTEs activated the wound epithelialization with complete healing. Patients GAIS results after rejuvenation program: Optimal cosmetic results 78.9%; significant improvement but not complete correction 9.7%; improvement but required additional correction 11.3%. The program significantly reduces the biological age and Frailty index that evidences about the decrease in risk of aging disease appearance.
Conclusion & Significance: 3D cultivation, in vivo modeling of SCs niche and prenatal SCs exosomes can significantly improve results of the use of SCs in regenerative medicine.