RESUMO
Considerable interest has been generated for the development of suitable corneal endothelial graft alternatives through cell-tissue engineering, which can potentially alleviate the shortage of corneal transplant material. The advent of less invasive suture-less key-hole surgery options such as Descemet's Stripping Endothelial Keratoplasty (DSEK) and Descemet's Membrane Endothelial Keratoplasty (DMEK), which involve transplantation of solely the endothelial layer instead of full thickness cornea, provide further impetus for the development of alternative endothelial grafts for clinical applications. A major challenge for this endeavor is the lack of specific markers for this cell type. To identify genes that reliably mark corneal endothelial cells (CECs) in vivo and in vitro, we performed RNA-sequencing on freshly isolated human CECs (from both young and old donors), CEC cultures, and corneal stroma. Gene expression of these corneal cell types was also compared to that of other human tissue types. Based on high throughput comparative gene expression analysis, we identified a panel of markers that are: i) highly expressed in CECs from both young donors and old donors; ii) expressed in CECs in vivo and in vitro; and iii) not expressed in corneal stroma keratocytes and the activated corneal stroma fibroblasts. These were SLC4A11, COL8A2 and CYYR1. The use of this panel of genes in combination reliably ascertains the identity of the CEC cell type.
Assuntos
Proteínas de Transporte de Ânions/genética , Antiporters/genética , Colágeno Tipo VIII/genética , Células Endoteliais/metabolismo , Endotélio Corneano/metabolismo , Expressão Gênica , Proteínas de Membrana/genética , Adulto , Idoso , Proteínas de Transporte de Ânions/metabolismo , Antiporters/metabolismo , Autopsia , Biomarcadores/metabolismo , Colágeno Tipo VIII/metabolismo , Ceratócitos da Córnea/citologia , Ceratócitos da Córnea/metabolismo , Substância Própria/citologia , Substância Própria/metabolismo , Células Endoteliais/citologia , Endotélio Corneano/citologia , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Especificidade de Órgãos , Cultura Primária de CélulasRESUMO
MicroRNAs (miRNAs) are important endogenous regulators of gene expression. The specific regulation at both the transcription and the translation level (inhibition or mRNA degradation) opens an avenue to use these small RNA molecules as potential targets for the development of novel drugs as well as for the diagnosis of several human diseases. Important information about the role of a miRNA in disease can be deduced by mimicking or inhibiting its activity and examining its impact on the phenotype/behaviour of the cell or organism. Modulating the activity of a miRNA is expected to lead to improvement in disease symptoms and this implies that the target miRNA plays an important role in the disease. It is also now possible to develop miRNA-based therapeutic products that can either increase or decrease the levels of proteins in pathophysiological conditions such as cancer, cardiovascular diseases, viral diseases, metabolic disorders and programmed cell death. The commercial potential of miRNA and related drugs is expected to exponentially increase within the next few years, yet there are several areas in miRNA biology and delivery that need to be extensively investigated.