Mental disorder and the outcome of HIV/AIDS in low-income and middle-income countries: a systematic review.

OBJECTIVES: To conduct a systematic review of the literature to examine the interrelationship between mental health and treatment outcomes in low-income and middle-income settings; to update the work of Collins et al. (2006). DESIGN: Systematic review of peer-reviewed articles that examined one of the following: the effects of mental disorders (including cognitive impairment) upon engagement with treatment and/or adherence; their influence upon HIV-related clinical outcomes; and the impact of interventions for mental disorder. METHODS: Articles about mental health and HIV/AIDS were included if they were published after 2005 and addressed one of the areas of interest described above. Systematic methods were used for searching, screening, and data extraction. Studies employing quantitative measures of exposures and outcomes wherein all participants had a diagnosis of HIV/AIDS were included. RESULTS: This review found ample and moderately consistent evidence that adverse mental health and alcohol consumption are associated with reduced adherence. Variation in measurement and the relative paucity of work meant that interpretation of studies examining engagement with care and other clinical outcomes was difficult. Evidence on the efficacy and effectiveness of mental health interventions in low-income and middle-income settings was very limited. CONCLUSION: This review suggests that psychosocial factors, namely, depression and alcohol may have adverse effects upon HIV-related outcomes. However, further large, high-quality studies examining outcomes other than adherence are needed. There is also an urgent need for randomized controlled trials of interventions for mental disorder and a need to investigate their impact upon HIV-related outcomes

Abnormal RNA Stability in Amyotrophic Lateral Sclerosis

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) share key features, including accumulation of the RNA-binding protein TDP-43. TDP-43 regulates RNA homeostasis, but it remains unclear whether RNA stability is affected in these disorders. We use Bru-seq and BruChase-seq to assess genome-wide RNA stability in ALS patient-derived cells, demonstrating profound destabilization of ribosomal and mitochondrial transcripts. This pattern is recapitulated by TDP-43 overexpression, suggesting a primary role for TDP-43 in RNA destabilization, and in postmortem samples from ALS and FTD patients. Proteomics and functional studies illustrate corresponding reductions in mitochondrial components and compensatory increases in protein synthesis. Collectively, these observations suggest that TDP-43 deposition leads to targeted RNA instability in ALS and FTD, and may ultimately cause cell death by disrupting energy production and protein synthesis pathways

Inhibition of PIP4Kγ ameliorates the pathological effects of mutant huntingtin protein.

The discovery of the causative gene for Huntington's disease (HD) has promoted numerous efforts to uncover cellular pathways that lower levels of mutant huntingtin protein (mHtt) and potentially forestall the appearance of HD-related neurological defects. Using a cell-based model of pathogenic huntingtin expression, we identified a class of compounds that protect cells through selective inhibition of a lipid kinase, PIP4Kγ. Pharmacological inhibition or knock-down of PIP4Kγ modulates the equilibrium between phosphatidylinositide (PI) species within the cell and increases basal autophagy, reducing the total amount of mHtt protein in human patient fibroblasts and aggregates in neurons. In two Drosophila models of Huntington's disease, genetic knockdown of PIP4K ameliorated neuronal dysfunction and degeneration as assessed using motor performance and retinal degeneration assays respectively. Together, these results suggest that PIP4Kγ is a druggable target whose inhibition enhances productive autophagy and mHtt proteolysis, revealing a useful pharmacological point of intervention for the treatment of Huntington's disease, and potentially for other neurodegenerative disorders

Ubiquilin-1 overexpression increases the lifespan and delays accumulation of Huntingtin aggregates in the R6/2 mouse model of Huntington's disease.

Huntington's Disease (HD) is a neurodegenerative disorder that is caused by abnormal expansion of a polyglutamine tract in huntingtin (htt) protein. The expansion leads to increased htt aggregation and toxicity. Factors that aid in the clearance of mutant huntingtin proteins should relieve the toxicity. We previously demonstrated that overexpression of ubiqulin-1, which facilitates protein clearance through the proteasome and autophagy pathways, reduces huntingtin aggregates and toxicity in mammalian cell and invertebrate models of HD. Here we tested whether overexpression of ubiquilin-1 delays or prevents neurodegeneration in R6/2 mice, a well-established model of HD. We generated transgenic mice overexpressing human ubiquilin-1 driven by the neuron-specific Thy1.2 promoter. Immunoblotting and immunohistochemistry revealed robust and widespread overexpression of ubiquilin-1 in the brains of the transgenic mice. Similar analysis of R6/2 animals revealed that ubiquilin is localized in huntingtin aggregates and that ubiquilin levels decrease progressively to 30% during the end-stage of disease. We crossed our ubiquilin-1 transgenic line with R6/2 mice to assess whether restoration of ubiquilin levels would delay HD symptoms and pathology. In the double transgenic progeny, ubiquilin levels were fully restored, and this correlated with a 20% increase in lifespan and a reduction in htt inclusions in the hippocampus and cortex. Furthermore, immunoblots indicated that endoplasmic reticulum stress response that is elevated in the hippocampus of R6/2 animals was attenuated by ubiquilin-1 overexpression. However, ubiquilin-1 overexpression neither altered the load of htt aggregates in the striatum nor improved motor impairments in the mice

Development and feasibility testing of an integrated PTSD and adherence intervention cognitive processing therapy-life steps (CPT-L) to improve HIV outcomes: Trial protocol

Despite high rates of Post-Traumatic Stress Disorder (PTSD) in persons living with HIV (PLWH) and poor HIV-related health outcomes associated with PTSD, an effective evidence-based treatment for PTSD symptoms in PLWH does not exist. Negative reinforcement conceptual models posit that avoidant behavior (hallmark symptom of PTSD) demonstrated by PLWH with co-occurring PTSD can contribute to poor antiretroviral therapy (ART) adherence. However, research evaluating the impact of evidence-based treatment for PTSD among HIV infected populations on HIV outcomes is scarce. The Cognitive Processing Therapy (CPT) protocol is an evidence-based PTSD treatment that may address internalized stigma with targeted modifications and improve ART adherence and subsequent viral suppression through reduction of avoidant coping. This study will be the first pilot open-label randomized control trial (RCT) to test feasibility of an integrated evidence-based PTSD treatment (CPT) with an adherence intervention (Lifesteps) delivered in a Ryan White clinic to improve PTSD symptoms, adherence to ART, and retention in HIV care. Primary aims are to (1) conduct theater testing of the CPT and Lifesteps research protocol and evaluate acceptability (n = 12) and (2) deliver a modified CPT protocol (CPT-Lifesteps, or CPT-L) in 60 PLWH/PTSD exploring impact of CPT-L on PTSD symptoms and HIV outcomes compared to a Lifesteps + Standard of Care condition. This innovative research extends PTSD treatment approaches as a paradigm to reduce barriers to ART adherence. Findings of this innovative study are significant because they support the Undetectable = Untransmittable (UU) campaign and can help prevent the transmission of HIV infection through increased viral suppression

Restoration of ubiquilin levels in animals carrying the R6/2 transgene and accompanying changes in htt protein levels.

<p>(A) Immunoblots of lysates made from dissected brain regions of 9 week-old mice with different genotypes obtained after crossing UBQ1 transgenic mice with R6/2 mice and probed for ubiquilin (upper panel) and for actin (lower panel). Note successful overexpression of ubiquilin in animals carrying the ubiquilin-1 transgene. (B) Immunoblot analysis similar to A, but this time showing ubiquilin levels in different regions of the brain (Ce  =  cerebellum, BS  =  Brain stem, FC  =  frontal cortex, St  =  striatum, Hi  =  hippocampus, VC =  visual cortex). Note overexpression of ubiquilin in all tissues of transgenic mice carrying the ubiquilin-1 transgene. (C) Immunoblots of equal amounts of protein in lysates made from the striatum (St), hippocampus (Hi) and frontal cortex (FC) of 9 week-old mice from the same cross mentioned above (W = non-transgenic for UBQ1 or R6/6, U = UBQ1 transgenic, R = R6/2 transgenic, R/U = R6/2-UBQ1 double transgenic) and probed for ubiquilin or actin. (D) Immunoblots of brain lysates from end-stage mice showing R6/2-UBQ1 double transgenic mice have higher ubiquilin levels detected in the resolving gel than R6/2 animals, which correlated with decreased ubiquilin that was trapped in aggregates in the stacking gel. Also included is a lysate from a 19-week old WT animal (right lane). (E and F) Quantification of ubiquilin levels in 9-week (E) and end-stage or equivalent time-point animals (F) showing the amount of ubiquilin protein expression in animals with different genotypes used in our study. Note that ubiquilin levels are reduced by 70% in end-stage R6/2 animals compared to wild type age-matched controls (<i>p</i> = 0.0011). Furthermore, ubiquilin levels were fully restored in the brains of end-stage R6/2-UBQ1 double transgenic mice compared to R6/2 transgenic mice (<i>p</i> = 0.0017). (G) Immunoblot of same lysates shown in D with EM48 antibody showing ubiquilin overexpression in R6/2-UBQ1 double transgenic mice have reduced levels of soluble mutant htt protein accumulation compared to R6/2 animals. Note the WT lysate was loaded on the left lane. (H) Quantification of soluble htt protein shown in panel G.</p

Ubiquilin is found in htt inclusions and its levels decrease in the R6/2 model of HD.

<p>(A) Confocal staining of a 15-week old end-stage R6/2 brain section of the hippocampus showing staining with anti-ubiquilin (red), anti-htt EM48 (green) and DAPI (blue). The merged image of the red and green fluorescence images shows ubiquilin colocalizes with htt inclusions (arrows). Bar = 15 µm. (B) Equal amounts of protein in whole brain homogenates from three end-stage R6/2 mice and three 19 week-old WT mice were immunoblotted for ubiquilin and for actin. Note the decline in ubiquilin levels in end-stage R6/2 animals.</p

Generation of transgenic mice that overexpress human ubiquilin-1.

<p>(A) Schematic of the Thy1.2 expression construct used to generate ubiquilin-1 transgenic mice. Human ubiquilin-1 with an N-terminal FLAG tag was cloned in the appropriate orientation between the XhoI site of the Thy1.2 expression cassette. (B) Southern Blot of the first generation offspring of two founder mice (48 and 62). (C) Validation of a PCR genotyping protocol. Amplification of the transgene was only observed in mice that Southern blotting revealed to be positive. (D) Immunoblots of brain cortical lysates with an anti-FLAG antibody and for tubulin indicated that line 62 offspring express higher levels of FLAG-ubiquilin-1 than line 48. (E) Immunoblots of equal amounts of total brain lysates from 12 month-old WT mouse, 12 month-old Ubqln-1 48 transgenic mouse, 12 month-old Ubqln-1 62 transgenic mouse and end stage 15 week-old R6/2 transgenic mouse. The top panel was probed with a monoclonal anti-ubiquilin antibody (Invitrogen antibody clone 3D5E2) and the lower panel with a different monoclonal anti-ubiquilin antibody (Novus antibody clone 5F5). Note two immunoreactive ubiquilin bands are seen at ∼70 kDa and at ∼90 kDa, which we presume is a modified form of ubiquilin. Both blots were also probed for actin to ensure equal loading. (F) Cryostat sections of a Ubqln-1 62 transgenic mouse brain (a–f) and WT mouse brain (g-i) showing anti-FLAG antibody staining (Alexa 594, left panels) and corresponding DAPI staining (center panels) and the result of merging the fluorescent and DAPI signals (right hand panels). The brain sections shown are of the hippocampus (a–c and g–i) and cerebellum (d–f). Identical exposure settings were used for the left hand panels.</p

Ubiquilin-1 overexpression modifies aggregate load in the hippocampus and cortex but not the striatum.

<p>(A) Representative fluorescence microscopy images of EM48 and DAPI stained cryostat sections of the CA1 region of the hippocampus in R6/2 transgenic and R6/2-UBQ1 double transgenic mouse at 6 weeks, 9 weeks and following end-stage euthanasia. Bar = 15 µm. (B) Similar to A, but showing representative sections from the dentate gyrus in end-stage mice. Bar = 15 µm. (C) Quantification of htt inclusions >0.5 µm in size in the CA1 region of the hippocampus at 6 weeks, 9 weeks and end-stage R6/2 and R6/2-UBQ1 double transgenic mice. The R6/2-UBQ1 double transgenic mice contained 22% fewer inclusions than R6/2 mice at 6 weeks (<i>p = </i>0.04), but not at the other times. (D) Quantification of htt inclusions >1 µm in size in the CA1 region of the hippocampus at 6 weeks, 9 weeks and end-stage R6/2 and R6/2-UBQ1 double transgenic mice. The R6/2-UBQ1 double transgenic mice had 40% fewer inclusions at 9 weeks compared to R6/2 transgenic mice (<i>p</i> = 0.027). (E and F) Similar to B and C, but showing htt inclusions in the cortex. R6/2-UBQ1 double transgenic mice had 8.5% fewer inclusions greater than 0.5 µm at the end-stage of disease. (G, H) Similar to E and F, but comparing inclusions in the striatum. There was no difference in the number of inclusions in the striatum between the two genotypes at any time point.</p