Abstract
Retinal degeneration is considered as a condition ensued by different blinding disorders such as retinitis pigmentosa, age-related macular degeneration, and diabetic retinopathy, which can cause loss of photoreceptor cells and also lead to significant vision deficiencies. Although there is no efficient treatment in this domain, transplantation of stem cells has been regarded as a therapeutic approach for retinal degeneration. Thus, the purpose of this study was to analyze the potential of human bone marrow-derived mesenchymal stem cells (hBMSCs) to differentiate into photoreceptor cells via transfection of microRNA (miRNA) in vitro for regenerative medicine purposes. To this end, miR-183/96/182 cluster was transfected into hBMSCs; then, qRT-PCR was performed to measure the expression levels of miR-183/96/182 cluster and some retina-specific neuronal genes such as OTX2, NRL, PKCα, and recoverin. CRX and rhodopsin (RHO) levels were also measured through qRT-PCR and immunocytochemistry, and subsequently, cellular change morphology was detected. The findings showed no changes in the morphology of the given cells, and the expression of the neuroretinal genes such as OTX2, NRL, and PKCα. Moreover, recoverin was upregulated upon miR-183/-96/-182 overexpression in cultured hBMSCs. Ectopic overexpression of the miR-183 cluster could further increase the expression of CRX and RHO at the messenger RNA (mRNA) and protein levels. Furthermore, the data indicated that the miR-183 cluster could serve as a crucial function in photoreceptor cell differentiation. In fact, miRNAs could be assumed as potential targets to exploit silent neuronal differentiation. Ultimately, it was suggested that in vitro overexpression of miR-183 cluster could trigger reprogramming of the hBMSCs to retinal neuron fate, especially photoreceptor cells.
Highlights
Degenerations and dystrophies in various neural retina subtypes are known as the major causes of eye disorders leading to blindness [1]
In order to prove the stemness of the bone marrow mesenchymal stem cells (BMSCs) derived from human bone marrow, the cells were analyzed against specific BMSCs antibodies and flow cytometric analysis showed positive morphological features as well as qualitative properties of the BMSCs. e population of BMSCs displayed positive expression of CD34 marker (Figure 2)
MiR-183 cluster was transfected into the human bone marrow-derived mesenchymal stem cells (hBMSCs). e results showed that increased expression levels of miR-183 cluster could lead to a rising trend in the expression of some photoreceptor markers. ese genes had a key role in the differentiation of photoreceptors
Summary
Degenerations and dystrophies in various neural retina subtypes are known as the major causes of eye disorders leading to blindness [1]. Ese findings have prompted researchers to attempt their induction into photoreceptors for the treatment of degenerative retinal diseases [1] In this domain, the bone marrow is acknowledged as an ideal source of pluripotent stem cells that has a highly undifferentiated and selfrenewing potential. It was found that treatment of mimic-miR-183/96/182 could induce photoreceptor-like cell differentiation in hBMSCs through upregulating orthodenticle homeobox 2 (OTX2) and cone-rod homeobox (CRX), transcription factor neural retina leucine zipper (NRL), and protein kinase C (PKC). Is study indicated that overexpression of the miR-183/96/182 in hBMSCs was a useful tool for direct photoreceptor differentiation via regulating multiple key photoreceptor-specific genes; it was suggested as a therapeutic potential for retinal degenerative diseases The BMSC-derived photoreceptor-like cells expressed recoverin and RHO membrane proteins in mature photoreceptors. is study indicated that overexpression of the miR-183/96/182 in hBMSCs was a useful tool for direct photoreceptor differentiation via regulating multiple key photoreceptor-specific genes; it was suggested as a therapeutic potential for retinal degenerative diseases
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