HTT Research

Transgene experiments

Alongside widespread vector supply and transgene expression, we demonstrate that a successful human mutant HTT mRNA and huntingtin protein lowering in the mind of a large-animal model of HD.

Although the sample size in this analysis was limited, we observed a tendency toward a dose-dependent reduction of huntingtin that was human following AAV5-miHTT administration.

BDNF genetics

Interest in compounds acting on a transcriptional system which supports the transcription of the BDNF gene (among others) is sustained by the fact that positive outcomes have been observed in HD versions after raising BDNF levels by way of transplanting cells engineered to overexpress BDNF to a compound model of HD or bringing BDNF through gene use.

Genetics of Huntington

Transgenic mouse research

Transgenic overexpression of BDNF from the cerebral cortex can delay the development of disease phenotypes from the R6/1 HD mouse model carrying the very first 63 amino acids of mutant human huntingtin protein and the genetic reduction of BDNF in precisely the same R6/1 mice triggers a more rapid phenotyping.

Mice lacking BDNF show a hind limb clasping phenotype similar to that found in mouse models of HD.

Cystamine boosts the HD phenotype in mice by enhancing release from the Golgi compartment at mind cells.

The RNAseq count level data of the human huntington gene knocking mouse model with varying CAG repeats changes in expression levels between mouse striatum samples and homozygous for the 20 CAG repeat allele (Q20), representing wild-type, and those heterozygous for the 175 CAG repeat allele (Q175), demonstrating HD pathology, were queried utilizing lmFit in the limma package.

HTT CAG repeat in various cell types, we analyzed whether CAG-correlated gene-expression signature in lymphoblastoid cell lines can also be enriched in genes associated with HTT CAG repeat length in human HD post-mortem brain.

DiFiglia et al. reported that an improvement in engine functions of R6/2 mice expressing N-terminal mutant Htt fragments following a single intrastriatal injection of cholesterol conjugated siRNA 21 At a rat model for HD, Franich et al. determined the shift in motor functions and equilibrium as a symptomatic mark for the expression of mutant Htt in rat brains.

Research in animal models are determining efficacy of remedies or to affirm disease pathogenesis. Huntington’s disease is an inherited autosomal dominant neurodegenerative disorder characterized by motor dysfunction, psychiatric disturbances, and innovative dementia 1, two HD is caused by an unstable CAG repeat expansion in the gene encoding huntingtin (HTT) on chromosome 4, resulting in an elongated polyglutamine (polyQ) elongate in the amino terminus of the HTT protein 3 ; the disorder is therefore related to a mutant form of the HTT protein that contains 36 or more glutamine residues. PGC-1α represents another important player in the link between mHTT, transcriptional dysregulation, and adrenal dysfunction ( Johri et al., 2013). PGC-1α is a transcriptional co-activator that modulates the manifestation of nuclear-encoded mitochondrial genes and modulates several metabolic processes, such as mitochondrial biogenesis and oxidative phosphorylation ( Wu et al., 1999 ; Puigserver and Spiegelman, 2003).

PGC-1α null mice manifest HD-like features including, striatal neuronal loss, hypothermia and motor alterations ( Weydt et al., 2006; Lucas et al., 2012 ). The term of PGC-1α is significantly downregulated in MSNs compared to other striatal cells in HD patients and transgenic mouse models ( Cui et al., 2006 ; Weydt et al., 2006 ). PGC-1α expression impairment in HD is expected, at least in part, to the disturbance of mHTT with all the CREB/TAF4 signaling pathway ( Cui et al., 2006 ), that can be regarded as the major regulator of PGC-1α expression ( Herzig et al., 2001).


Chromatin immunoprecipitation analysis conducted in murine striatal-like cells derived from WT (STHdhQ7) and HD (STHdhQ111) mice didn’t reveal differences in CREB/TAF4 binding into the PGC-1α promoter between both cell types ( Cui et al., 2006 ) suggesting that additional mechanisms may be included in PGC-1α expression handicap (discussed elsewhere in this review).

How P53 influences Huntington?

P53 protein levels and activity are induced in the brain of HD patients and in mouse and cell versions of HD33, explaining at least in part, the low tumor incidence observed in HD patients ( Sørensen et al., 1999 ; Bae et al., 2005). MHTT closely interacts with p53, also it has been proposed that such interaction disrupts the recruitment of the E3 ligase Mdm2, thus increasing p53 stabilization ( Steffan et al., 2000 ; Bae et al., 2005 ). Up-regulation of p53 contributes to induced expression of distinct mitochondria related proteins (e.g., Bax and Puma, linked to mitochondrial depolarization) and activation of apoptosis ( Chipuk et al., 2004; La Spada and Morrison, 2005). The function of p53 in mediating mitochondrial dysfunction in HD has been confirmed when principal neurons expressing mHTT were treated using the p53 inhibitor pifithrin-α and demonstrated enhanced mitochondrial membrane potential (MMP) and increased cell viability ( Bae et al., 2005 ). Lately, p53 was shown to also take part in mediating mitochondrial related necrosis and fragmentation in HD via direct interaction with mitochondrial fission protein Drp1 (dynamin-related protein; Guo et al., 2013, 2014).

The mechanism by which p53 inhibition exerts neuroprotection is poorly known. In peripheral tissues (lymphoblast, myoblast and fibroblasts) mitochondria pose an enlarged morphology, whereas neurons are characterized by enhanced adrenal fragmentation ( Panov et al., 2002 ; Squitieri et al., 2006, 2010; Kim et al., 2010; Jin et al., 2013). Altered mitochondrial structure correlates with mitochondrial dysfunction in all HD cells, that’s triggered by decreased electron transport chain activity, oxygen consumption, Ca2+ loading and decreased ATP and NAD+ generation ( Oliveira, 2010).


It’s been suggested that parasitic HTT (mHTT)-mediated adrenal abnormalities significantly influence MSNs on account of the high-energy demand of the neuronal subtype ( Ferrante et al., 1991; Pickrell et al., 2011). This is one hypothesis that explains the increased vulnerability of MSNs in HD ( Ferrante et al., 1991; Mitchell and Griffiths, 2003). In support of the hypothesis, mitochondria isolated from the striatum of adult rats demonstrated higher sensitivity to Ca2+ triggered membrane permeabilization than mitochondria from the cerebral cortex, suggesting that striatal neurons are selectively vulnerable to metabolic stress ( Brustovetsky et al., 2003).

Cell-selective neuropathology include; cell-type-specific processing or localization of mHTT ( Li et al., 2000 ; Menalled et al., 2002 ), abnormal interactions between mHTT and brain region specific protein spouses and tissue specific differences in CAG instability ( Kennedy et al., 2003 ; Goula et al., 2012 ). All these processes play important roles and they’re not the subject of this review, though they may also contribute to raise mitochondrial stress.

Pcr Arrays

These gene sets were compared with the human HD brain gene-expression information from Sage Bionetworks (; cerebellum, prefrontal cortex and visual cortex) from samples for that we had previously determined HTT CAG allele size, using a similar algorithm as found in the sigPathway. SignArray from Anygenes have a specific pcr primer array for Huntington and Alzheimer.

Significantly, gene accession numbers were used to match lymphoblast and human brain expression information, and correlation between gene expression and CAG repeat length in the individual brain gene-expression data were calculated. By genotype-phenotype research in HTT CAG allelic series designed to check the full range of ordinary and HD-associated allele sizes in human lymphoblastoid cell lines, it has emerged that the constant relationship between CAG dimensions and dominant biochemical steps outcome extends beneath the enlarged CAG HD scope.

The view of the HTT CAG repeat as a functional polymorphism with dominant quantitative biological effects that become overpowering at HD disease-producing CAG lengths can also be consistent with the demonstrated continuous impact of the CAG repeat encoded polyglutamine tract about the activity of huntingtin protein, a ubiquitously expressed early HEAT domain shown to be involved in a wide array of cellular processes.

What is Huntington disease? (overview)

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder brought on by a polyglutamine (polyQ) expansion in the huntingtin (HTT) protein. HTT is localized in the synapse, where it binds to proteins involved in neurotransmitter transport, in addition to pre- and postsynaptic organization and function, indicating that it serves as an important regulator of neurological activity. In contrast, mutant HTT impairs synaptic function in HD.

This defect occurs early in the absence of illustrated neuron loss, and is most likely the source of emotional, cognitive, learning, and memory symptoms in HD sufferers. A key factor in this decline is the progressive loss of excitatory corticostriatal glutamatergic input to medium-sized spiny neurons (MSNs), which has been associated with cell-autonomous and neuronal network mechanics.

Since the discovery of the genetic basis for the disease, mutant huntingtin (that), at 1993 ( MacDonald et al., 1993), there has been considerable effort toward creating therapeutic approaches for HD, and several compounds show beneficial effects in several HD cell and transgenic mouse models ( Li et al., 2005; Ray and Shoulson, 2011 ; Guo et al., 2013). However, human trials in HD are time consuming because of the slow progression of the disease, its insidious onset, and patient-to-patient variability ( MacDonald et al., 1993; Weir et al., 2011 ). There is also a need to incorporate a large cohort of patients since most of the clinical assessments are absolutely subjective (e.g., psychiatric tests) and also an inability to biopsy of the affected cells, nerves in the brain. Huntington’s disease (HD) is a deadly autosomal-dominant neurodegenerative disease caused by an enlarged trinucleotide CAG replicate in the gene encoding the huntingtin protein ( MacDonald et al., 1993). HD is a progressive disease that affects middle-age carriers, and also the intensity of the disease correlates with the length of the CAG repeat ( Lee et al., 2015). Patients affected by HD show a loss of neurons predominantly from the striatum and cortex that is progressively accompanied by a loss of voluntary and involuntary movements as well as cognitive and psychiatric disturbances.

HTT stem cell research

We evaluated this idea that has a small isogenic panel of heterozygous mouse Hdh CAG knock-in embryonic stem cell lines, containing just four alleles drawn from the normal (CAG 18) and expanded (CAG 48, CAG 89, CAG 109) human ranges. The results confirmed a constant approach did efficiently distinguish genes whose level of expression was changed in a way that was significantly correlated with CAG allele size. Notably, these transcriptional changes invisibly biological pathways/processes associated with Huntington functional pathways also shown the CAG-correlated genes formed a distinct and relatively small set of molecular responses that were not identified using the traditional dichotomous Htt CAG enlarged allele versus Htt wild-type allele contrast. While encouraging, the identification of a CAG-correlated transcriptional response from this otherwise isogenic allelic cell panel could not reveal whether the strategy will be sufficiently powerful to distinguish the effects of the CAG repeat in panels of individual cells or tissues where, in addition to a variety of different environmental and possibility factors, the genetic history is highly variable from person to person. HSF1 plays a fundamental role in HD pathogenesis (recently reviewed by Gomez-Pastor et al., 2017b ). Studies at which HSF1 null mice were crossbred with all the R6/2 mice demonstrated that deficiency of HSF1 worsens neurodegeneration and disease progression ( Hayashida et al., 2010) whilst HD transgenic mice overexpressing a constitutive active form of HSF1 significantly ameliorated HD symptoms ( Fujimoto et al., 2005 ). The levels of HSF1 and its activity are strongly depleted in the striatum of patients with HD and in cell and mouse models of HD ( Hay et al., 2004; Labbadia et al., 2011 ; Chafekar and Duennwald, 2012; Riva et al., 2012; Maheshwari et al., 2014 ; Gomez-Pastor et al., 2017a). HSF1 depletion is caused by improper up-regulation of MSNs in two proteins, including the Protein Kinase CK2α’ and E3 ligase Fbxw7, which phosphorylate and ubiquitylate HSF1, respectively, signaling the protein for proteasomal degradation ( Gomez-Pastor et al., 2017a ). It is considered that decreased levels and activity of HSF1 contribute to neuronal dysfunction and pathogenesis, implying HSF1 as a potential therapeutic target for HD intervention ( Sittler et al., 2001; Neef et al., 2011 ). This hypothesis is supported by CK2α’ allele knock-out studies in the HD KIQ175 mouse model, which resulted in increased HSF1 levels and neuronal chaperone expression, rescued MSN’s morphology and synapse formation, and ameliorated weight reduction associated with HD ( Gomez-Pastor et al., 2017a ). Huntington’s disease (HD) is a neurodegenerative disease brought on by an expanded CAG repeat in the huntingtin (HTT) gene, causing the protein to misfold and aggregate.