THE REGULATION OF PROTEINS 14-3-3 AND THE HIPPO PATHWAY AFFECT THE ADIPOGENESIS OF 3T3-L1

LIM, G.; UHART, M.; DEL VELIZ, S.; BUSTOS, D. M.

Salta

Conferencia; Joint LV AnnualSAIB Meeting and XIV PABMB Congress; 2019

SAIB-PABMB

Manytranscription factors act sequentially to activate adipocytedifferentiation. Multiple signal transduction pathways govern theadipocyte physiology. The Hippo kinase pathway is the main regulatorof proliferation and differentiation of stem cells and adipocyteprecursor cells. The 14-3-3 protein family, comprised of 7 paralogsin mammals, interacts with components of the Hippo kinase pathwayregulating the adipogenic differentiation, although the specificityof these proteins in the process remains elusive. Deciphering thesubcellular dynamics of 14-3-3 proteins will be instrumental in thediscovery of new targets for the manipulation of stem cell fatedecisions or treatment of chronic diseases, such as obesity and type2 diabetes.Invitro adipocyte differentiation occurs by the addition of anAdipogenic Differentiation Medium (ADM) including Dulbecco´s ModifiedEagle Medium, 10% Fetal Bovine Serum, synthetic drugs (dexamethasone,IBMX, rosiglitazone) and peptide hormones (insulin). Since thesementioned chemical drugs are not present in physiologicalenvironments, it would be important to achieve a correct activatedadipogenic program using fewer drugs and with natural origin. We havedecided to evaluate the adipogenic potential of glucagon-like peptide1 (GLP-1) analogs. In contrast to Native GLP-1 action, which israpidly degraded by circulating Dipeptidyl Peptidase-4, GLP-1 analogshave shown positive effects on glycemic control and body weight dueto their greater stability and longer half-life.Inthis project, experiments were performed on 3T3-L1 preadipocytes.First, the effect of different combination of drugs for 7 days inadipogenesis was evaluated. Then, we carried out qPCR experiments tomeasure the gene expression of 14-3-3 and the most important proteinsof the Hippo pathway, on days 3 and 7 of differentiation.Wehave determined that conditions resulting in greater adipogenicdifferentiation showed higher levels of Hippo pathway proteins and14-3-3 gamma and beta isoforms on day 7. These effects were speciallyevident when IBMX was replaced by GLP-1 in the ADM. These resultssuggest that different inducers (glucocorticoids, thiazolidinediones,incretins), have different abilities for regulating the expression of14-3-3 and hippo pathway proteins, thus affecting adipogenicdifferentiation. We are currently focused on elucidating themechanisms of action of these drugs to have a greater understandingof the events that are necessary for adipogenesis to occur.p { margin-bottom: 0.25cm; direction: ltr; font-variant: normal; color: rgb(0, 0, 10); letter-spacing: normal; line-height: 120%; text-align: left; break-inside: auto; text-decoration: none; break-before: auto; break-after: avoid; }p.western { font-family: "Times New Roman", serif; font-size: 12pt; font-style: normal; font-weight: normal; }p.cjk { font-family: "Arial Unicode MS"; font-size: 12pt; font-style: normal; font-weight: normal; }p.ctl { font-family: "Times New Roman"; font-size: 12pt; font-style: normal; font-weight: normal; }a:link { color: rgb(0, 0, 10); text-decoration: underline; }