Study of the links between the dysregulations of metabolism and epigenetics marks in Huntington’s disease

We want to focus on epigenetic dysregulation in Huntington’s Disease (HD), a pathogenic mechanism implicated in accelerated aging of striatal neurons. Specifically, we will investigate the interplay between altered energy metabolism and epigenetic impairment in HD striatal neurons to identify new targets/pathways for disease-modifying intervention. We aim to obtain detailed maps of histone post-translational modifications (PTMs), especially of methylations, acetylation and the recently described lactylation, which might be critical in the HD brain. Indeed, these PTMs are tightly regulated by the metabolic status of the cells. We will use proteomics which is the best suited approach to identify and quantify multiple protein PTMs. We consider working on the striatum of WT, R6/1 transgenic mice and the more progressive Q140 knock in model at various stages of disease, to assess evolution of histone PTMs and metabolism with aging. Additionally, to get a dynamic view of the links between metabolic and epigenetic imbalance in HD, we will inject intraperitoneally HD mice and controls with 13C-glucose and analyze over a time course the incorporation of 13C into histone PTMs. Finally, acetyl-CoA, the precursor for histone lysine acetylation, has been shown to be locally produced in the nucleus, by either acetyl-CoA synthetase 2 (ACSS2), ATP-citrate lyase (ACLY) or the pyruvate dehydrogenase complex. Regarding lactylation, it is currently unknown where, and by which enzymes, the pool of lactate used for modifying histone lysines by lactylation is produced. ACSS2 is a very good candidate, as it can catalyze the production of acyl-CoA molecules from the corresponding short chain fatty acids (SCFA). To address the question of the production of metabolites in the vicinity of chromatin in striatal cells, we will use epigenomics (ChIPseq or CUT&tag) to get the genomic distribution of ACSS2 and ACLY and compare it to distributions of acetyl and lactyl histone marks.

Towards a detailed understanding of the regulation of gene expression by acetylation and lactylation of histone proteins

In eukaryotic cells, DNA is wrapped around histone proteins to form chromatin. Dynamic modification of histones by various chemical structures enables fine regulation of gene expression. Alterations in these complex regulatory mechanisms are at the root of many diseases. Histone lysine acetylation is known to induce gene expression. Other structures can be added to histones, whose effects on transcription remain largely to be elucidated. Most of them, like lactylation discovered in 2019, depend on cellular metabolism. We have begun to study lactylation in the context of murine spermatogenesis. This process of cellular differentiation is a model of choice for studying the regulation of transcription, due to the dramatic changes in chromatin composition and the gene expression program. We have generated novel epigenetic profiles consisting of the genome-wide distribution of acetylated and lactylated marks on three histone H3 lysines. The aim of this thesis is to contribute to the deciphering of the “histone code”, firstly by studying the role of lactylations on the transcriptional program. Secondly, the prediction of chromatin states will be refined by integrating our new data with existing epigenomic data at the two studied cellular stages, within neural network models.

ROLE OF UNFOLDED PROTEIN RESPONSE IN MAINTAINING THE SPERMATOGONIAL STEM CELL POOL IN THE ADULT MOUSE

Adverse conditions (oxidative stress, imbalanced lipid, glucose or calcium levels, or inflammation) induce the accumulation of abnormal proteins resulting in ER stress. The Unfolded Stress Response (UPR) is activated to restore cellular homeostasis, but severe or chronic stress results in apoptotic cell death. Uncontrolled UPR signaling promotes many human diseases (diabetes, Parkinson's, Alzheimer's, liver disease, cancer...), but nothing is known about its implication in adult male sterility. Spermatozoa production relies on Spermatogonial Stem Cells (SSC) which are maintained by self-renewal throughout life. We have shown that the clonogenic activity of SSC is drastically impaired after ER stress through differentiation entry. An HTS screen has highlighted 2 of the 3 UPR branches as being involved in the clonogenic activity of SSC in vitro. The role of these 2 UPR pathways will be further investigated in SSC cultures of mice to determine whether they are involved in the induction of cell death or in the balance between self renewal and differentiation. In treated SSC cultures, cell death, cell cycle, induction of differentiation and synergy between UPR pathways will be analyzed. As the effect of each pathway is mediated by transcriptional factors, the target genes will be characterized by RNAseq in order to identify the gene networks controlled by UPR effectors and involved in the fate of SSC. For the most relevant pathway, an in vivo study will confirm the role of the UPR effector in CSS property.

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