Premature termination codon readthrough upregulates progranulin expression and improves lysosomal function in preclinical models of GRN deficiency

BACKGROUND: Frontotemporal lobar degeneration (FTLD) is a devastating and progressive disorder, and a common cause of early onset dementia. Progranulin (PGRN) haploinsufficiency due to autosomal dominant mutations in the progranulin gene (GRN) is an important cause of FTLD (FTLD-GRN), and nearly a quarter of these genetic cases are due to a nonsense mutation. Premature termination codons (PTC) can be therapeutically targeted by compounds allowing readthrough, and aminoglycoside antibiotics are known to be potent PTC readthrough drugs. Restoring endogenous PGRN through PTC readthrough has not previously been explored as a therapeutic intervention in FTLD. METHODS: We studied whether the aminoglycoside G418 could increase PGRN expression in HEK293 and human induced pluripotent stem cell (hiPSC)-derived neurons bearing the heterozygous S116X, R418X, and R493X pathogenic GRN nonsense mutations. We further tested a novel substituted phthalimide PTC readthrough enhancer in combination with G418 in our cellular models. We next generated a homozygous R493X knock-in hiPSC isogenic line (R493X(-/-) KI), assessing whether combination treatment in hiPSC-derived neurons and astrocytes could increase PGRN and ameliorate lysosomal dysfunction relevant to FTLD-GRN. To provide in vivo proof-of-concept of our approach, we measured brain PGRN after intracerebroventricular administration of G418 in mice expressing the V5-tagged GRN nonsense mutation R493X. RESULTS: The R418X and R493X mutant GRN cell lines responded to PTC readthrough with G418, and treatments increased PGRN levels in R493X(-/-) KI hiPSC-derived neurons and astrocytes. Combining G418 with a PTC readthrough enhancer increased PGRN levels over G418 treatment alone in vitro. PGRN deficiency has been shown to impair lysosomal function, and the mature form of the lysosomal protease cathepsin D is overexpressed in R493X(-/-) KI neurons. Increasing PGRN through G418-mediated PTC readthrough normalized this abnormal lysosomal phenotype in R493X(-/-) KI neuronal cultures. A single intracerebroventricular injection of G418 induced GRN PTC readthrough in 6-week-old AAV-GRN-R493X-V5 mice. CONCLUSIONS: Taken together, our findings suggest that PTC readthrough may be a potential therapeutic strategy for FTLD caused by GRN nonsense mutations

World-Wide FINGERS Network: A global approach to risk reduction and prevention of dementia

© 2020 The Authors. Alzheimer\u27s & Dementia published by Wiley Periodicals, Inc. on behalf of Alzheimer\u27s Association Reducing the risk of dementia can halt the worldwide increase of affected people. The multifactorial and heterogeneous nature of late-onset dementia, including Alzheimer\u27s disease (AD), indicates a potential impact of multidomain lifestyle interventions on risk reduction. The positive results of the landmark multidomain Finnish Geriatric Intervention Study to Prevent Cognitive Impairment and Disability (FINGER) support such an approach. The World-Wide FINGERS (WW-FINGERS), launched in 2017 and including over 25 countries, is the first global network of multidomain lifestyle intervention trials for dementia risk reduction and prevention. WW-FINGERS aims to adapt, test, and optimize the FINGER model to reduce risk across the spectrum of cognitive decline—from at-risk asymptomatic states to early symptomatic stages—in different geographical, cultural, and economic settings. WW-FINGERS aims to harmonize and adapt multidomain interventions across various countries and settings, to facilitate data sharing and analysis across studies, and to promote international joint initiatives to identify globally implementable and effective preventive strategies

Samspillsentreprisers påvirkning på konfliktnivået i norsk byggebransje

Norsk byggebransje preges av et høyt konfliktnivå. Dette medfører store økonomiske og ressursmessige tap for både aktørene i bransjen og samfunnet generelt. For en bransje som utgjør en betydelig del av norsk næringsliv, er det viktig å utarbeide tiltak som kan motvirke det høye konfliktnivået. I en samspillsveileder, reklameres det for at det blir færre konflikter i samspillsentrepriser. Denne oppgaven undersøker i hvilken grad samspillsentrepriser faktisk fungerer som konfliktforebyggende tiltak i norsk byggebransje. Studien er gjennomført som en kombinasjon av de to kvalitative forskningsmetodene: litteraturstudie og intervjuer. I litteraturstudien er tidligere forskning innhentet og analysert. De kvalitative intervjuene er gjennomført med totalt 21 relevante fagpersoner fra aktørgruppene entreprenør, byggherre, advokater og andre (forskere og rådgivere). Funnene fra denne studien viser at konflikter oppstår som følge av at usikkerhet, utydelighet eller feil sørger for at to parter med ulike hensikter og mål blir uenige, uten å finne en rask felles løsning. Samspillsentrepriser vil motvirke disse årsakene i form av at samspillselementet tidlig involvering bidrar til økt forståelse, færre utydeligheter og mindre behov for endringer utover i prosjektet. Samtidig vil samspill kunne bidra til en økonomisk stabilitet og integrerte prosjektteam, hvor aktørene jobber mot et felles mål. Totalt sett kan svært mange aspekter med samspillsenterpriser fungere konfliktreduserende. Samtlige intervjuobjekter påstod også at samspillsentrepriser er konfliktdempende, men da gitt at prosjektet og aktørene har de riktige forutsetningene for å gjennomføre. Både teorien og intervjuene tilsier at samspillsentreprise er en krevende prosess å gjennomføre, og det kreves riktig innstilling, tilstrekkelig tid og kompetanse. Dersom forutsetningene ikke er på plass, vil effektene av samspillet potensielt endres. 45 prosent av intervjuobjektene mener at samspillsentrepriser kan være konfliktøkende, dersom prosjektet og aktørene ikke innfrir forutsetningene. De fleste aktører er kapable til å utvikle en samspillskontrakt, selv om den potensielt kan bli ubalansert. Gjennom studien, viser det seg at en ikke kan bestemme seg for å inngå et samspillsprosjekt, og forvente gevinster. Det krever innsats for å opparbeide og vedlikeholde samspillskulturen. I hvilken grad samspillsentrepriser er konfliktreduserende er helt avhengig av samspillskulturen. Derfor konkluderes det med at en samspillsentreprise i seg selv ikke nødvendigvis er konfliktforebyggende, men det kan legge til rette for en god samspillskultur, som utvilsomt vil være konfliktdempende. Nøkkelord: Konflikthåndtering, konfliktårsaker, samspill, byggebransj

Neuropathological and behavioral characterization of aged Grn R493X progranulin-deficient frontotemporal dementia knockin mice

Abstract Frontotemporal lobar degeneration (FTLD) causes a spectrum of clinical presentations of frontotemporal dementia (FTD), including progressive changes in behavior, personality, executive function, and language. Up to 20% of familial FTLD cases are caused by progranulin (GRN) haploinsufficiency (FTD-GRN), with one of the most common causal variant being a nonsense mutation at arginine 493 (R493X). Recently, a genetic knockin FTD-GRN mouse model was generated bearing this GrnR493X mutation, at the analogous arginine in murine Grn. Aged, homozygous GrnR493X mice (GrnR493X/R493X) have been shown to phenotypically replicate several neuropathological hallmarks previously demonstrated in Grn null mice. We conducted a comprehensive neuropathological and behavioral assessment of 18 month old GrnR493X/R493X mice, observing a striking lysosomal dysfunction and thalamic neurodegeneration not previously described in this model, as well as a male-specific increase in generalized anxiety. These findings provide additional phenotypic markers of pathogenesis in aged GrnR493X/R493X mice that will contribute to better defining mechanisms underlying FTD-GRN, and offer relevant outcome measures for preclinical efficacy testing of novel therapeutics that target nonsense mutations leading to this devastating disease.Medicine, Faculty ofOther UBCMedicine, Department ofNeurology, Division ofReviewedFacult

Sleep and its regulation: An emerging pathogenic and treatment frontier in Alzheimer's disease.

A majority of patients with Alzheimer's disease (AD) experience some form of sleep disruption, including nocturnal sleep fragmentation, increased daytime napping, decreased slow-wave sleep (SWS, stage N3), and decreased rapid-eye-movement sleep (REM). Clinical studies are investigating whether such sleep disturbances are a consequence of the underlying disease, and whether they also contribute to the clinical and pathological manifestations of AD. Emerging research has provided a direct link between several of these sleep disruptions and AD pathophysiology, suggesting that treating sleep disorders in this population may target basic mechanisms of the disease. Here, we provide a comprehensive review of sleep disturbances associated with the spectrum of AD, ranging from the preclinical stages through dementia. We discuss how sleep interacts with AD pathophysiology and, critically, whether sleep impairments can be targeted to modify the disease course in a subgroup of affected AD patients. Ultimately, larger studies that fully utilize new diagnostic and experimental tools will be required to better define the most relevant sleep disturbance to target in AD, the interventions that best modulate this target symptom, and whether successful early intervention can modify AD risk and prevent dementia

Delayed Amyloid Plaque Deposition and Behavioral Deficits in Outcrossed AβPP/PS1 Mice

Alzheimer’s disease (AD) is a progressive neurodegenerative dementia characterized by amyloid plaque accumulation, synapse/dendrite loss, and cognitive impairment. Transgenic mice expressing mutant forms of amyloid-β precursor protein (AβPP) and presenilin-1 (PS1) recapitulate several aspects of this disease and provide a useful model system for studying elements of AD progression. AβPP/PS1 mice have been previously shown to exhibit behavioral deficits and amyloid plaque deposition between 4–9 months of age. We crossed AβPP/PS1 animals with mice of a mixed genetic background (C57BL/6 × 129/SvJ) and investigated the development of AD-like features in the resulting outcrossed mice. The onset of memory-based behavioral impairment is delayed considerably in outcrossed AβPP/PS1 mice relative to inbred mice on a C57BL/6 background. While inbred AβPP/PS1 mice develop deficits in radial-arm water maze performance and novel object recognition as early as 8 months, outcrossed AβPP/PS1 mice do not display defects until 18 months. Within the forebrain, we find that inbred AβPP/PS1 mice have significantly higher amyloid plaque burden at 12 months than outcrossed AβPP/PS1 mice of the same age. Surprisingly, inbred AβPP/PS1 mice at 8 months have low plaque burden suggesting that plaque burden alone cannot explain the accompanying behavioral deficits. Analysis of AβPP processing revealed that elevated levels of soluble Aβ correlate with the degree of behavioral impairment in both strains. Taken together, these findings suggest that animal behavior, amyloid plaque deposition, and AβPP processing are sensitive to genetic differences between mouse strains