Deciphering transcriptional regulation of NF-kB target genes using integrative omics approaches

Descripción del proyecto

Análisis de los mecanismos de activación transcripcional de NF-kB

Los factores de transcripción que forman parte de la familia del factor nuclear-kB (NK-kB, por sus siglas en inglés) dirigen la expresión de genes fundamentales que están implicados en la respuesta inmunitaria, la proliferación y la apoptosis. Al mismo tiempo, nuevas evidencias indican que el NK-kB tiene una función en la aparición y la proliferación del cáncer. Por lo tanto, para desarrollar un tratamiento contra el NK-kB para el cáncer, sería necesario actuar de manera selectiva sobre los genes NK-kB asociados con la transformación maligna. Con esto en mente, el proyecto financiado con fondos europeos RegulatioNFkB se propone investigar el mecanismo de activación transcripcional mediado por el NF-kB centrándose en las regiones promotoras de los genes que responden al NK-kB. Los investigadores usarán la edición del genoma y la espectrometría de masas para identificar los factores de transcripción y otras proteínas que interactúan en los promotores de estos genes.

Objetivo

NF-kB is a family of transcription factors that regulate expression of genes that are involved in various cellular processes such as apoptosis, proliferation and differentiation. The NF-kB pathway is implicated in the initiation and progression of cancer and cancer cell drug resistance. Unfortunately, global NF-kB inhibition is harmful to the immune system; therefore, inhibition of a subset of NF-kB target genes that are linked to malignant transformation would be more appropriate. Different NF-kB target genes display distinct transcription activation kinetics, indicating that the molecular mechanisms driving expression of NF-kB target genes are strikingly diverse. Thus, it is highly interesting to study the factors that are involved in transcriptional activation of individual NF-kB responsive genes, both from a fundamental and clinical perspective. Here, we will develop two innovative strategies based on CRISPR/Cas9 based genome targeting combined with mass spectrometry-based technology to isolate single NF-kB target gene promoters upon NF-kB pathway activation and identify their constituents. We will also apply a complementary approach, ATAC-sequencing, to identify transcription factors that are involved in activation of NF-kB responsive genes. Moreover, we will study the underlying mechanisms and characterize the function of proteins that interact with NF-kB responsive promoters. Finally, the described mechanisms discovered in cancer cell lines will be verified and validated in small intestinal organoids, a non-transformed cell model for homeostasis in the intestinal epithelium. This ambitious and multidisciplinary project will greatly benefit from my expert knowledge regarding the NF-kB pathway combined with the expertise of the host laboratory in mass spectrometry and genomics. The project is expected to have a major impact in the research area of gene expression regulation in general and molecular cancer research focusing on the NF-kB pathway in particular.