27 research outputs found

    Altered H3 histone acetylation impairs high-fidelity DNA repair to promote cerebellar degeneration in spinocerebellar ataxia type 7.

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    A common mechanism in inherited ataxia is a vulnerability of DNA damage. Spinocerebellar ataxia type 7 (SCA7) is a CAG-polyglutamine-repeat disorder characterized by cerebellar and retinal degeneration. Polyglutamine-expanded ataxin-7 protein incorporates into STAGA co-activator complex and interferes with transcription by altering histone acetylation. We performed chromatic immunoprecipitation sequencing ChIP-seq on cerebellum from SCA7 mice and observed increased H3K9-promoter acetylation in DNA repair genes, resulting in increased expression. After detecting increased DNA damage in SCA7 cells, mouse primary cerebellar neurons, and patient stem-cell-derived neurons, we documented reduced homology-directed repair (HDR) and single-strand annealing (SSA). To evaluate repair at endogenous DNA in native chromosome context, we modified linear amplification-mediated high-throughput genome-wide translocation sequencing and found that DNA translocations are less frequent in SCA7 models, consistent with decreased HDR and SSA. Altered DNA repair function in SCA7 may predispose the subject to excessive DNA damage, leading to neuron demise and highlights DNA repair as a therapy target

    ATF6-mediated signaling contributes to PARP Inhibitor Resistance in Ovarian Cancer.

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    High grade serous ovarian cancer (HGSOC) is the deadliest ovarian cancer histotype due in-part to the lack of therapeutic options for chemotherapy resistant disease. Poly(ADP)-ribose polymerase inhibitors (PARPi) represent a targeted treatment. However, PARPi resistance is becoming a significant clinical challenge. There is an urgent need to overcome resistance mechanisms to extend disease-free intervals. We established isogeneic PARPi-sensitive and -resistant HGSOC cell lines. In three PARPi-resistant models there is a significant increase in AP-1 transcriptional activity and DNA repair capacity. Using RNA-seq and an shRNA screen, we identified Activating Transcription Factor 6 (ATF6) as a mediator of AP-1 activity, DNA damage response, and PARPi resistance. In publicly available datasets, ATF6 expression is elevated in HGSOC and portends a poorer recurrence free survival. In a cohort of primary HGSOC tumors, higher ATF6 expression significantly correlated to PARPi resistance. In PARPi-resistant cell lines and a PDX model, inhibition of a known ATF6 regulator, p38, attenuated AP-1 activity and RAD51 foci formation, enhanced DNA damage, significantly inhibited tumor burden, and reduced accumulation of nuclear ATF6. Implications: This study highlights that a novel p38-ATF6 mediated AP-1 signaling axis contributes to PARPi resistance and provides a clinical rationale for combining PARPi and AP-1 signaling inhibitors

    Haploinsufficiency networks identify targetable patterns of allelic deficiency in low mutation ovarian cancer.

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    Identification of specific oncogenic gene changes has enabled the modern generation of targeted cancer therapeutics. In high-grade serous ovarian cancer (OV), the bulk of genetic changes is not somatic point mutations, but rather somatic copy-number alterations (SCNAs). The impact of SCNAs on tumour biology remains poorly understood. Here we build haploinsufficiency network analyses to identify which SCNA patterns are most disruptive in OV. Of all KEGG pathways (N=187), autophagy is the most significantly disrupted by coincident gene deletions. Compared with 20 other cancer types, OV is most severely disrupted in autophagy and in compensatory proteostasis pathways. Network analysis prioritizes MAP1LC3B (LC3) and BECN1 as most impactful. Knockdown of LC3 and BECN1 expression confers sensitivity to cells undergoing autophagic stress independent of platinum resistance status. The results support the use of pathway network tools to evaluate how the copy-number landscape of a tumour may guide therapy

    Neuronal Ndst1 depletion accelerates prion protein clearance and slows neurodegeneration in prion infection.

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    Select prion diseases are characterized by widespread cerebral plaque-like deposits of amyloid fibrils enriched in heparan sulfate (HS), a abundant extracellular matrix component. HS facilitates fibril formation in vitro, yet how HS impacts fibrillar plaque growth within the brain is unclear. Here we found that prion-bound HS chains are highly sulfated, and that the sulfation is essential for accelerating prion conversion in vitro. Using conditional knockout mice to deplete the HS sulfation enzyme, Ndst1 (N-deacetylase / N-sulfotransferase) from neurons or astrocytes, we investigated how reducing HS sulfation impacts survival and prion aggregate distribution during a prion infection. Neuronal Ndst1-depleted mice survived longer and showed fewer and smaller parenchymal plaques, shorter fibrils, and increased vascular amyloid, consistent with enhanced aggregate transit toward perivascular drainage channels. The prolonged survival was strain-dependent, affecting mice infected with extracellular, plaque-forming, but not membrane bound, prions. Live PET imaging revealed rapid clearance of recombinant prion protein monomers into the CSF of neuronal Ndst1- deficient mice, neuronal, further suggesting that HS sulfate groups hinder transit of extracellular prion protein monomers. Our results directly show how a host cofactor slows the spread of prion protein through the extracellular space and identify an enzyme to target to facilitate aggregate clearance

    <i>Ndst1</i><sup><i>f/f</i></sup><i>SynCre-</i> and <i>SynCre+</i> mice infected with subfibrillar prions: survival times, brain lesions, and biochemical properties.

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    (A) Survival curves of SynCre- and SynCre+ mice intracerebrally inoculated with RML or ME7 prions (RML: n = 7 SynCre- and 8 SynCre+ mice; ME7: n = 16 SynCre- and 17 SynCre+ mice). (B) Brain sections (hippocampus) immunolabelled for PrPSc or GFAP or stained with hematoxylin and eosin (HE) show prion aggregate distribution and morphology, spongiform degeneration, and astrogliosis in SynCre- and SynCre+ brains. Scale bar = 50 μm. (C) Lesion profiles of RML- and ME7-infected SynCre- and SynCre+ mice (1-dorsal medulla, 2-cerebellum, 3-hypothalamus, 4-medial thalamus, 5-hippocampus, 6-septum, 7-cerebral cortex, 8- cerebral peduncle and 9- cerebellar peduncle) (RML: n = 5 mice per genotype; ME7: n = 6 SynCre- and 7 SynCre+ mice). (D) PrPSc immunoblots (post-PK digest) show electrophoretic mobility and glycoprofile of PK-digested PrPSc. N = 6 per genotype. (E) Immunolabelling for microglia (Iba1) (representative images of hippocampus) and (F) quantification of the Iba1-stained area in CC, CT, HP, TH. Scale bar = 50 μm. (G) Representative immunoblots for PrPSc (post PK digest) from 30 μg brain, and (H) quantification of PrPSc levels in the whole brain lysate at terminal disease. *P (TIF)</p
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