title: MicroRNA expression profiling of Leishmania donovani-infected host cells uncovers the regulatory role of MIR30A-3p in host autophagy.
abstract: Leishmania is an obligate intracellular parasite that replicates inside phagolysosomes or parasitophorous vacuoles (PV) of the monocyte/macrophage lineage. It reprograms macrophages and produces a metabolic state conducive to successful infection and multiplication. MicroRNAs (miRNAs), a class of small 22 to 24 nucleotide noncoding regulatory RNAs alter the gene expression and consequently proteome output by targeting mRNAs, may play a regulatory role in modulating host cell functions. In the present study, we demonstrate the novel regulatory role of host microRNA, MIR30A-3p in modulation of host cell macroautophagy/autophagy after infection with L. donovani. Our in vitro studies showed that MIR30A-3p expression was significantly enhanced after L. donovani infection in a time-dependent manner. Transient transfection with a MIR30A-3p inhibitor followed by L. donovani infection promoted the autophagic response and decreased the intracellular parasite burden in both THP-1 cells and human monocyte-derived macrophages (HsMDM). BECN1/Beclin 1, the mammalian ortholog of yeast Vps30/Atg6, is a key autophagy-promoting protein that plays a key role in the regulation of cell death and survival. We report BECN1-dependent modulation of host cell autophagy in response to L. donovani infection. Pretreatment of L. donovani-infected macrophages with the MIR30A-3p mimic decreased and with antagomir increased the expression of BECN1 protein. We demonstrate that BECN1 is a potential target of MIR30A-3p and this miRNA negatively regulates BECN1 expression. Our present study reveals for the first time a novel role of MIR30A-3p in regulating autophagy-mediated L. donovani elimination by targeting BECN1. The present study has significant impact for the treatment of visceral leishmaniasis.
authors.affiliation: a School of Life Sciences, Jawaharlal Nehru University , New Delhi , India.
authors.forename: Alok Kumar
authors.initials: AK
authors.lastname: Singh
authors.affiliation: b Cell Death and Differentiation Research Laboratory, National Institute of Immunology , New Delhi , India.
authors.forename: Rajeev Kumar
authors.initials: RK
authors.lastname: Pandey
authors.affiliation: b Cell Death and Differentiation Research Laboratory, National Institute of Immunology , New Delhi , India.
authors.forename: Chandrima
authors.initials: C
authors.lastname: Shaha
authors.affiliation: a School of Life Sciences, Jawaharlal Nehru University , New Delhi , India.
authors.forename: Rentala
authors.initials: R
authors.lastname: Madhubala
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics*; D000071183:Autophagy-Related Proteins / Q000378:metabolism; D000071186:Beclin-1 / Q000378:metabolism; D002460:Cell Line; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions / Q000235:genetics*; D006801:Humans; D007893:Leishmania donovani / Q000502:physiology*; D007898:Leishmaniasis, Visceral / Q000235:genetics* / Q000469:parasitology* / Q000473:pathology; D008264:Macrophages / Q000378:metabolism / Q000469:parasitology / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009000:Monocytes / Q000473:pathology; D010271:Parasites / Q000502:physiology; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics
chemical_list: D000071183:Autophagy-Related Proteins; D000071186:Beclin-1; C546086:MIRN30b microRNA, human; D035683:MicroRNAs; D044763:Ubiquitin-Conjugating Enzymes; C455621:ATG3 protein, human
keywords: BECN1; Leishmania donovani; MIR30A-3p; THP-1 cells; autophagy; gene expression; human monocyte-derived macrophages; microRNAs