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Analysis of the therapeutic potential of the human recombinant RSPO1 protein in regeneration of small intestine in an animal model using tissue Engeneering technologies

Grant number: 15/11128-3
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): September 01, 2015
Effective date (End): May 31, 2020
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Mari Cleide Sogayar
Grantee:Gabriel Levin
Home Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated scholarship(s):17/01072-6 - Analysis of the therapeutic potential of the human recombinant RSPO1 protein in regeneration of the small intestine in an animal model using tissue engineering technologies, BE.EP.DD


R-Spondin (RSPO) belongs to a family of proteins known for inducing the WNT pathway, which plays a pivotal and diversified role in cell proliferation, differentiation, migration and death during embryogenesis and, also, in the adult. RSPO genes are expressed in several signaling centers known for their critical roles during development, regulating diverse tissue-specific processes, such as bone formation, proliferation of pancreatic beta cells and intestinal stem cells, also being associated with several types of cancer. Inappropriate expression of RSPOs and WNT signaling may lead to several pathological conditions. The mitogenic activity of RSPO1 protein in intestinal stem cells may offer several therapeutic opportunities. R-Spondins are critical for maintenance of small intestine crypt stem cells niche. This project proposes to generate overproducing cell clones of the human recombinant RSPO1 in CHO-dhfr-/- cells using the pNU1 mammalian expression vector, to obtain a purified and biologically active protein to be used in intestinal regeneration. To evaluate the rhRSPO1 activity in tissue regeneration, RSPO1 will be used individually or in combination with other recombinant peptide growth factors, such as VEGF and PDGF-BB, in culture of murine intestinal organoid units using a Tissue Engineered Small Intestine (TESI) model in NOD/SCID receptor mice. The coding sequence of hRSPO1 gene has already been synthesized and sub-cloned into the pNU1 vector to be used for transfection into CHO-dhfr-/- cells. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MASELLI, KATHRYN M.; LEVIN, GABRIEL; GEE, KRISTIN M.; LEEFLANG, ELISABETH J.; CARREIRA, ANA CLAUDIA O.; SOGAYAR, MARI CLEIDE; GRIKSCHEIT, TRACY C. R-Spondin1 enhances wnt signaling and decreases weight loss in short bowel syndrome zebrafish. BIOCHEMISTRY AND BIOPHYSICS REPORTS, v. 25, MAR 2021. Web of Science Citations: 0.
LEVIN, GABRIEL; KOGA, BRUNA ANDRADE AGUIAR; BELCHIOR, GUSTAVO GROSS; CARREIRA, ANA CLAUDIA OLIVEIRA; SOGAYAR, MARI CLEIDE. Production, purification and characterization of recombinant human R-spondin1 (RSPO1) protein stably expressed in human HEK293 cells. BMC Biotechnology, v. 20, n. 1 JAN 20 2020. Web of Science Citations: 0.

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