Perturbed actin cap as a new personalized biomarker in primary fibroblasts of Huntington’s disease patients

Gharaba, Saja; Paz, Omri; Feld, Lea; Abashidze, Anastasia; Weinrab, Maydan; Muchtar, Noam; Baransi, Adam; Shalem, Aviv; Sprecher, Uri; Wolf, Lior; Wolfenson, Haguy; Weil, Miguel

doi:10.3389/fcell.2023.1013721

Cited by 5 publications

(5 citation statements)

References 44 publications

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“…Similar to exploring common variant associations, we are likely underpowered in any single disease category to identify significant associations. There have been many studies elucidating morphological features associated with various diseases, but they often contained larger sample sizes and incorporated more specialized cell types 22 , 65 , 66 . Extending our current study to diverse cell types and increasing the number of samples for clinical disease categories will be a critical next step in efforts to link cell morphology to human illnesses.…”

Section: Discussionmentioning

confidence: 99%

High-dimensional phenotyping to define the genetic basis of cellular morphology

Tegtmeyer,

Arora,

Asgari

et al. 2024

Nat Commun

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The morphology of cells is dynamic and mediated by genetic and environmental factors. Characterizing how genetic variation impacts cell morphology can provide an important link between disease association and cellular function. Here, we combine genomic sequencing and high-content imaging approaches on iPSCs from 297 unique donors to investigate the relationship between genetic variants and cellular morphology to map what we term cell morphological quantitative trait loci (cmQTLs). We identify novel associations between rare protein altering variants in WASF2, TSPAN15, and PRLR with several morphological traits related to cell shape, nucleic granularity, and mitochondrial distribution. Knockdown of these genes by CRISPRi confirms their role in cell morphology. Analysis of common variants yields one significant association and nominate over 300 variants with suggestive evidence (P < 10−6) of association with one or more morphology traits. We then use these data to make predictions about sample size requirements for increasing discovery in cellular genetic studies. We conclude that, similar to molecular phenotypes, morphological profiling can yield insight about the function of genes and variants.

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Section: Discussionmentioning

confidence: 99%

High-dimensional phenotyping to define the genetic basis of cellular morphology

Tegtmeyer,

Arora,

Asgari

et al. 2024

Nat Commun

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“…Over 100 interactions between huntingtin and different proteins are known [ 57 ]. The abnormal morphology of the HD fibroblast is correlated with actin cap deficiency, and it was suggested that this unique feature can be used as a personalized biomarker of HD [ 58 ]. The Rho-Rac GTPase cascade was identified to be involved in the reorganization of the cytoskeleton by actin-binding proteins cofilin and profilin, and it was concluded that the loss of phosphoSer138 in profilin is correlated with the symptomatic course of HD [ 59 ].…”

Section: Discussionmentioning

confidence: 99%

Actin Cytoskeleton Polymerization and Focal Adhesion as Important Factors in the Pathomechanism and Potential Targets of Mucopolysaccharidosis Treatment

Gaffke

Rintz

Pierzynowska

et al. 2023

Cells

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The main approach used in the current therapy of mucopolysaccharidosis (MPS) is to reduce the levels of glycosaminoglycans (GAGs) in cells, the deposits considered to be the main cause of the disease. Previous studies have revealed significant differences in the expression of genes encoding proteins involved in many processes, like those related to actin filaments, in MPS cells. Since the regulation of actin filaments is essential for the intracellular transport of specific molecules, the process which may affect the course of MPSs, the aim of this study was to evaluate the changes that occur in the actin cytoskeleton and focal adhesion in cells derived from patients with this disease, as well as in the MPS I mouse model, and to assess whether they could be potential therapeutic targets for different MPS types. Western-blotting, flow cytometry and transcriptomic analyses were employed to address these issues. The levels of the key proteins involved in the studied processes, before and after specific treatment, were assessed. We have also analyzed transcripts whose levels were significantly altered in MPS cells. We identified genes whose expressions were changed in the majority of MPS types and those with particularly highly altered expression. For the first time, significant changes in the expression of genes involved in the actin cytoskeleton structure/functions were revealed which may be considered as an additional element in the pathogenesis of MPSs. Our results suggest the possibility of using the actin cytoskeleton as a potential target in therapeutic approaches for this disease.

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“…SH-SY5Y cells comprise a neuroblastoma cell line and are commonly utilised in HD research [21,22]. Reduced cell proliferation and altered nuclear morphology have been noted in HD post mortem brains previously [25]. Furthermore, in HD patients, mutant and wild-type HTT proteins are found to be nuclear-localised, which is largely thought to be due to the propensity of mutant HTT to be highly promiscuous and bind various proteins, including the nuclear master regulator REST within the cytoplasm [32].…”

Section: Discussionmentioning

confidence: 99%

“…As changes in nuclear and cell morphology have been noted in HD [24,25], these same parameters were also investigated in circHTT(2-6) overexpression cell lines. This was achieved through high-content imaging using an Operetta CLS (Perkin Elmer).…”

Section: Cell Morphology Analysismentioning

confidence: 99%

Functional Characterisation of the Circular RNA, circHTT(2-6), in Huntington’s Disease

Gantley

Stringer

Conn

et al. 2023

Cells

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Trinucleotide repeat disorders comprise ~20 severe, inherited, human neuromuscular and neurodegenerative disorders, which result from an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease (HD), results from expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Since non-coding RNAs have been implicated in the initiation and progression of many diseases, herein we focused on a circular RNA (circRNA) molecule arising from non-canonical splicing (backsplicing) of HTT pre-mRNA. The most abundant circRNA from HTT, circHTT(2-6), was found to be more highly expressed in the frontal cortex of HD patients, compared with healthy controls, and positively correlated with CAG repeat tract length. Furthermore, the mouse orthologue (mmu_circHTT(2-6)) was found to be enriched within the brain and specifically the striatum, a region enriched for medium spiny neurons that are preferentially lost in HD. Transgenic overexpression of circHTT(2-6) in two human cell lines—SH-SY5Y and HEK293—reduced cell proliferation and nuclear size without affecting cell cycle progression or cellular size, or altering the CAG repeat region length within HTT. CircHTT(2-6) overexpression did not alter total HTT protein levels, but reduced its nuclear localisation. As these phenotypic and genotypic changes resemble those observed in HD patients, our results suggest that circHTT(2-6) may play a functional role in the pathophysiology of this disease.

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Perturbed actin cap as a new personalized biomarker in primary fibroblasts of Huntington’s disease patients

Cited by 5 publications

References 44 publications

High-dimensional phenotyping to define the genetic basis of cellular morphology

High-dimensional phenotyping to define the genetic basis of cellular morphology

Actin Cytoskeleton Polymerization and Focal Adhesion as Important Factors in the Pathomechanism and Potential Targets of Mucopolysaccharidosis Treatment

Functional Characterisation of the Circular RNA, circHTT(2-6), in Huntington’s Disease

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