Proteomic Analysis of Huntington’s Disease Medium Spiny Neurons Identifies Alterations in Lipid Droplets

Tshilenge, Kizito-Tshitoko; Aguirre, Carlos Galicia; Bons, Joanna; Gerencser, Akos A.; Basisty, Nathan; Song, Sicheng; Rose, Jacob; Lopez-Ramirez, Alejandro; Naphade, Swati; Loureiro, Ashley; Battistoni, Elena; Milani, Mateus; Wehrfritz, Cameron; Holtz, Anja; Hetz, Claudio; Mooney, Sean D.; Schilling, Birgit; Ellerby, Lisa M.

doi:10.1016/j.mcpro.2023.100534

Cited by 13 publications

(6 citation statements)

References 140 publications

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“…Next, we sought to assess whether RNA alterations were reflected at the protein level in developing HD72 MSNs. We compared our RNAseq data with our recently published proteomics dataset from developing HD72 and C116 MSNs [38]. We identified a considerable correlation between gene fold change values at both the RNA and protein levels (Pearson's r = 0.75, p-value < 2.2e-16), highlighting the potential impact of these alterations on cellular function (Supplemental Fig.…”

Section: Resultsmentioning

confidence: 99%

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Transcriptomic Characterization Reveals Disrupted Medium Spiny Neuron Trajectories in Huntington’s Disease and Possible Therapeutic Avenues

Aquirre

Tshilenge

Battistoni

et al. 2023

Preprint

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Huntington's disease (HD) is a neurodegenerative disorder caused by an expansion of CAG repeats in exon 1 of the HTT gene, ultimately resulting in the generation of a mutant HTT (mHTT) protein. Although mHTT is expressed in various tissues, it significantly affects medium spiny neurons (MSNs) in the striatum, resulting in their loss and subsequent motor function impairment in HD. While HD symptoms typically emerge in midlife, disrupted MSN neurodevelopment has an important role. To explore the effects of mHTT on MSN development, we differentiated HD induced pluripotent stem cells (iPSC) and isogenic controls into neuronal stem cells, and then generated a developing MSN population encompassing early, intermediate progenitors, and mature MSNs. Single-cell RNA sequencing revealed that the developmental trajectory of MSNs in our model closely emulated the trajectory of fetal striatal neurons. However, in the HD MSN cultures, the differentiation process downregulated several crucial genes required for proper MSN maturation, including Achaete-scute homolog 1 and members of the DLX family of transcription factors. Our analysis also uncovered a progressive dysregulation of multiple HD-related pathways as the MSNs matured, including the NRF2-mediated oxidative stress response and mitogen-activated protein kinase signaling. Using the transcriptional profile of developing HD MSNs, we searched the L1000 dataset for small molecules that induce the opposite gene expression pattern. Our analysis pinpointed numerous small molecules with known benefits in HD models, as well as previously untested novel molecules. A top novel candidate, Cerulenin, partially restored the DARPP-32 levels and electrical activity in HD MSNs, and also modulated genes involved in multiple HD-related pathways.

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Section: Resultsmentioning

confidence: 99%

“…NSCs and MSNs were differentiated as described in [38]. Briefly, HD72 and C116 iPSCs were turned into cell aggregates and driven towards a neuroepithelial fate that produced neural rosettes.…”

Section: Experimental Procedures Nsc and Developing Msn Differentiationmentioning

confidence: 99%

Transcriptomic Characterization Reveals Disrupted Medium Spiny Neuron Trajectories in Huntington’s Disease and Possible Therapeutic Avenues

Aquirre

Tshilenge

Battistoni

et al. 2023

Preprint

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“…Mutation of HTT leads to many changes in molecular processes that, although initially subtle, have already been observed in HD patient-derived iPSCs or embryonic stem cells (ESCs) [56][57][58][59]. At later steps of cell differentiation, these changes lead to disruptions that have significant implications for the functions of neural cells [31,56,[60][61][62][63][64][65]. Stem cell-derived neural lines are models that are widely used in HD research to investigate aspects of transcriptional dysregulation [63,66,67], but the underlying mechanisms appear to be diverse and complex due to the interplay of the factors involved [26].…”

Section: Discussionmentioning

confidence: 99%

Huntingtin loss-of-function contributes to transcriptional deregulation in Huntington’s disease

Emilia,

Agata,

Grażyna

et al. 2024

Preprint

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Huntington's disease (HD) is a fatal neurodegenerative disorder that is caused by the expansion of CAG repeats in the HTT gene, which results in a long polyglutamine (polyQ) tract in the huntingtin protein (HTT). In this study, we searched for networks of deregulated RNAs that contribute to initial transcriptional changes in HD neuronal cells and HTT-deficient cells. We used RNA-seq (including small RNA sequencing) to analyze a set of isogenic, human induced pluripotent stem cell (iPSC)-derived neural stem cells (NSCs); and we observed numerous changes in gene expression and substantial dysregulation of miRNA expression in HD and HTT-knockout (HTT-KO) cell lines. The gene set that was upregulated in both HD and HTT-KO cells was enriched in genes that are associated with DNA binding and regulation of transcription. For both of these models, we confirmed the substantial upregulation of the transcription factors (TFs) TWIST1, SIX1, TBX1, TBX15, MSX2, MEOX2 and FOXD1 in NSCs and medium spiny neuron (MSN)-like cells. Moreover, we identified miRNAs that were consistently deregulated in HD and HTT-KO NSCs and MSN-like cells, including miR-214, miR-199, and miR-9. We suggest that these miRNAs function in the network that regulates TWIST1 and HTT expression via regulatory feed-forward loop (FFL) in HD. Additionally, we reported that the expression of selected TFs and miRNAs tended to progressively change during the neural differentiation of HD cells, what was not observed in HTT-KO model. Based on comparing the HD and HTT-KO cell lines, we propose that early transcriptional deregulation in HD is largely caused by loss of HTT function.

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“…Recent studies showed that VPS13A is localized at sites where the endoplasmic reticulum (ER) and mitochondria are in close contact to enable lipid transfer required for mitochondria and lipid-droplet-related processes in cell lines ( Yeshaw et al, 2019 ). Interestingly, structural and functional changes in the ER-mitochondria contact sites leading to mitochondrial dysfunction ( Shirendeb et al, 2011 ; Cherubini et al, 2020 ), and aberrant lipid homeostasis or calcium signaling ( Aditi et al, 2016 ; Pchitskaya et al, 2018 ; Tshilenge et al, 2023 ) have been reported to contribute to the specific degeneration of the striatum in HD ( Browne, 2008 ).…”

Section: Discussionmentioning

confidence: 99%

Preserved VPS13A distribution and expression in Huntington’s disease: divergent mechanisms of action for similar movement disorders?

García-García,

Carreras-Caballé,

Coll-Manzano

et al. 2024

Front. Neurosci.

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VPS13A disease and Huntington’s disease (HD) are two basal ganglia disorders that may be difficult to distinguish clinically because they have similar symptoms, neuropathological features, and cellular dysfunctions with selective degeneration of the medium spiny neurons of the striatum. However, their etiology is different. VPS13A disease is caused by a mutation in the VPS13A gene leading to a lack of protein in the cells, while HD is due to an expansion of CAG repeat in the huntingtin (Htt) gene, leading to aberrant accumulation of mutant Htt. Considering the similarities of both diseases regarding the selective degeneration of striatal medium spiny neurons, the involvement of VPS13A in the molecular mechanisms of HD pathophysiology cannot be discarded. We analyzed the VPS13A distribution in the striatum, cortex, hippocampus, and cerebellum of a transgenic mouse model of HD. We also quantified the VPS13A levels in the human cortex and putamen nucleus; and compared data on mutant Htt-induced changes in VPS13A expression from differential expression datasets. We found that VPS13A brain distribution or expression was unaltered in most situations with a decrease in the putamen of HD patients and small mRNA changes in the striatum and cerebellum of HD mice. We concluded that the selective susceptibility of the striatum in VPS13A disease and HD may be a consequence of disturbances in different cellular processes with convergent molecular mechanisms already to be elucidated.

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Proteomic Analysis of Huntington’s Disease Medium Spiny Neurons Identifies Alterations in Lipid Droplets

Cited by 13 publications

References 140 publications

Transcriptomic Characterization Reveals Disrupted Medium Spiny Neuron Trajectories in Huntington’s Disease and Possible Therapeutic Avenues

Transcriptomic Characterization Reveals Disrupted Medium Spiny Neuron Trajectories in Huntington’s Disease and Possible Therapeutic Avenues

Huntingtin loss-of-function contributes to transcriptional deregulation in Huntington’s disease

Preserved VPS13A distribution and expression in Huntington’s disease: divergent mechanisms of action for similar movement disorders?

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