Abstract: FR-PO978
Role of ApoL1-miR193a Axis in Sox2-Mediated Reprogramming of Differentiated Podocytes
Session Information
- Pathology and Lab Medicine: Basic
November 08, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Pathology and Lab Medicine
- 1601 Pathology and Lab Medicine: Basic
Authors
- Jha, Alok, Feinstein Institute for Medical Research, Manhasset, New York, United States
- Ayasolla, Kamesh R., Feinstein Institute for Medical Research, Manhasset, New York, United States
- Kumar, Vinod, Feinstein Institute for Medical Research, Manhasset, New York, United States
- Vashistha, Himanshu, Ochsner Health System, New Orleans, Louisiana, United States
- Lan, Xiqian, Feinstein Institute for Medical Research, Manhasset, New York, United States
- Qayyum, Maleeha, Northwell health , Jamaica, New York, United States
- Chinnapaka, Sushma, Northwell health , Jamaica, New York, United States
- Adnani, Harsha, Feinstein Institute of Research, Glen Oaks, New York, United States
- Malhotra, Ashwani, Feinstein Institute Medical Research and NSLIJ, Manhasset, New York, United States
- Skorecki, Karl, Rambam Health Care Campus, Haifa, Israel
- Singhal, Pravin C., North Shore LIJ Health System, Great Neck, New York, United States
Background
Enhanced autophagy maintains molecular phenotype in differentiated podocytes. The Sox2 is known to initiate autophagy by repressing Mammalin Target of Rapamycin (mTOR) expression; however, Sox2-induced autophagy induction carries negative feedback on reprogramming of differentiation. Since miR193a suppresses Sox2 expression, it would modulate the Sox2-induced transient autophagy during an early step in reprogramming to pluripotency/differentiation. Because APOL1 inversely regulates miR193a, it would de-repress Sox2 and accelerate autophagy in differentiated podocytes
Methods
Human podocytes expressing vector, APOL1G0/G1/G2 were differentiated; protein blots were probed for Sox2, APOL1, nephrin, CD2AP, and GAPDH. RNAs were assayed for miR193a and cDNA amplified for APOL1, and Sox2. The silico method was used to analyze motifs on ApoL1 and its variants mRNA. MEME suite for motif identification, JASPARv2010 and STAMP tool for alignment, and database matching for identified motifs were used. 3D models of ApoL1 and its variants mRNA segments (with mutations and deletions) were generated. Structural model of Sox2 (template-based method Itasser) docking approach was used to form the ApoL1 and its variants RNA and Sox2 complexes. The thermodynamic properties of ApoL1G0, G1, and G2 mRNAs and Sox2 were analyzed.
Results
Differentiated G0-podocytes displayed enhanced Sox2 but decreased expression of miR193a; in contrast, G1- and G2-podocytes showed the opposite outcome. The MEME suite identified motifs on ApoL1, and its variants and the JASPARv2010 and STAMP database for motif matching suggested that Sox2 can bind on ApoL1G2 mRNA. The thermodynamic properties of RNA-protein interaction interface suggested that the ApoL1G2 mRNA and Sox2 form a very strong and stable complex with surface area 4443.8 Å2, solvation free energy gain upon the formation of the interface (ΔiG) -93.8 kcal/mol and free energy of assembly dissociation (ΔGdiss) 95.6 kcal/mol. The ApoL1G2 mRNA-Sox2 complex has 38 hydrogen bonding interactions.
Conclusion
The Sox2 binding with ApoL1 and its variants mRNA suggests that Sox2 has an RNA binding property. This interaction carries the potential to modulate Apol1-miR193a-induced downstream signaling.
Funding
- NIDDK Support