Genetic screens for heterochronic mutants in C. elegans, first reported twenty-eight years ago, defined an evolutionarily conserved pathway controlling growth and developmental timing in all bilateral organisms, and from which emerged the discovery of microRNAs. We discovered a fundamental biochemical mechanism that relates two mammalian homologues of heterochronic worm genes: the RNA binding protein LIN28 inhibits the biogenesis of let-7 microRNAs (miRNAs). We and others have implicated LIN28/let-7 in a sweeping range of biology including mammalian growth and developmental timing of sexual maturation, somatic cell reprogramming and pluripotency, germ cell development, cancer, inflammation, glycolytic metabolism, and diabetes. Our recent data connects the LIN28/let-7 axis to the shift towards glycolytic metabolism that accompanies reprogramming of somatic cells to pluripotency. We have also linked LIN28/let-7 to the distinctive amino acid metabolism of embryonic and pluripotent cells, in which flux through Threonine/ Methionine influences levels of s-adenosyl methionine, histone H3K4 methylation, and pluripotency. The ancestral C. elegans LIN28 gene exists in mammals as two highly related paralogs, A and B, which show different temporal and spatial patterns of tissue expression, different protein structures and modifications, and different RNA targets. This proposal will analyze transcriptional regulation, structure/function relationships, post- translational modifications, and effects on the proteome of the two paralogs in the context of somatic cell reprogramming and metabolism, testing the hypothesis that LIN28 confers a glycolytic metabolic state characteristic of embryonic and pluripotent cells. Illuminating the LIN28/let-7 pathway will provide fundamental insights into reprogramming while also shedding light on disease processes like cancer and diabetes. Understanding its mechanisms is destined to have major significance for biomedicine.
This project investigates the function of a recently described pathway involving the LIN28 RNA binding protein and one of its key targets, the let-7 family of microRNAs. LIN28/let-7 has been implicated in several aspects of human physiology and disease, including the regulation of growth, puberty, metabolism, cancer, and diabetes. Moreover, LIN28 is one of the factors that facilitates the resetting of specialized adult cells back to a state resembling embryonic stem cells, a process termed reprogramming. This application will investigate the mechanism by which LIN28/let-7 influences cell proliferation and metabolism, and will use reprogramming as an accessible in vitro model system to dissect the structure-function relationships of LIN28B and its targets. Insights into how LIN28 is regulated and the impact on its many targets may point towards improved methods to generate stem cells and novel therapeutic interventions in conditions like cancer and diabetes
|Kumar, Roshan M; Cahan, Patrick; Shalek, Alex K et al. (2014) Deconstructing transcriptional heterogeneity in pluripotent stem cells. Nature 516:56-61|
|Nguyen, Liem H; Robinton, Daisy A; Seligson, Marc T et al. (2014) Lin28b is sufficient to drive liver cancer and necessary for its maintenance in murine models. Cancer Cell 26:248-61|