Abstract Background Parkinson’s disease (PD) is a prevalent neurodegenerative disorder characterized by progressive motor dysfunction and broad cellular impairment, including significant disruptions in lysosomal function, lipid metabolism, and intracellular trafficking. Glycosphingolipids (GSLs), critical for various cellular processes, depend on effective lysosomal degradation. Aberrant GSL metabolism has been linked to PD pathology, and glycoprotein non-metastatic melanoma protein B (GPNMB) has emerged as a biomarker associated with lysosomal dysfunction and lipid imbalance in PD Objectives To assess the relationship between GPNMB and GSL levels in cerebrospinal fluid (CSF) and plasma from PD patients and controls within the BioFIND cohort. We also investigated potential sex differences and associations with PD-related biomarkers such as α-synuclein Methods GSL species and GPNMB protein levels were quantified using high-performance liquid chromatography (HPLC) and ELISA assays, respectively, in matched CSF and plasma samples from PD patients and controls Results Levels of the paraglobosides GSL species, alpha-2,3SpG and pGb were significantly elevated in the plasma of PD patients compared to healthy controls, while levels of the ganglioside GD1a and the lacto-series GSL, Le b combined (GD1a + Le b ), were significantly reduced in PD. GPNMB levels positively correlated with several GSL species in both plasma and CSF. Plasma GSLs and GPNMB concentrations were significantly higher in females compared to males, independent of PD diagnosis. CSF GPNMB correlated positively with age and α-synuclein concentrations Interpretation Our findings confirm that GSL metabolism is altered in PD. They also highlight significant sex-based biochemical variations in GSL and GPNMB levels, emphasizing the need for sex-specific analyses in PD biomarker research. The relationship between GSLs and GPNMB supports their potential as interconnected biomarkers of lipid pathology in PD.

Apolipoprotein E (ApoE) variants are central to Alzheimer’s disease (AD), Lewy body dementia (LBD) and Niemann-Pick disease type C (NPC). The ApoE4 variant elevates AD risk by 3-15-fold. ApoE’s normal function in lipid transport is known. The question remains how different ApoE isoforms cause cellular pathogenesis. We determined the effects of ApoE isoforms on lipid accumulation induced by inhibiting the endo-lysosomal cholesterol transporter NPC1. In human fibroblasts and astrocytes, NPC1 inhibition caused a 4-fold cholesterol accumulation and mis-localization with altered cholesterol sensing and increased synthesis of cholesterol and triglycerides. Total APP, APP C-terminal fragments (CTF) and BACE1 levels increased 3-fold. Remarkably, the intracellular neutral lipids co-localized with APP and APP C-terminal fragments. ApoE2 and ApoE3, but not ApoE4, reduced intracellular cholesterol levels by 67% and 62%, respectively, normalized APP, BACE, CTF, and improved cell survival. ApoE4 combined with a synthetic lipopeptide, which increased the proportion of large lipidated ApoE4 particles, corrected these abnormalities. This highlights ApoE in lipid pathogenesis and targeting ApoE4 lipidation to restore ApoE4 function.

Tightly regulated and highly adaptive lipid metabolic and transport pathways are critical to maintaining brain cellular lipid homeostasis and responding to lipid and inflammatory stress to preserve brain function and health. Deficits in the lipid handling genes APOE and GBA1 are the most significant genetic risk factors for Lewy body dementia and related dementia syndromes. Parkinson’s disease patients who carry both APOE4 and GBA1 variants have accelerated cognitive decline compared to single variant carriers. To investigate functional interactions between brain ApoE and GBA1, in vivo GBA1 inhibition was tested in WT versus ApoE-deficient mice. The experiments demonstrated glycolipid stress caused by GBA1 inhibition in WT mice induced ApoE expression in several brain regions associated with movement and dementia disorders. The absence of ApoE in ApoE-KO mice amplified complement C1q elevations, reactive microgliosis and astrocytosis after glycolipid stress. Mechanistically, GBA1 inhibition triggered increases in cell surface and intracellular lipid transporters ABCA1 and NPC1, respectively. Interestingly, the absence of NPC1 in mice also triggered elevations of brain ApoE levels. These new data show that brain ApoE, GBA1 and NPC1 functions are interconnected in vivo, and that the removal or reduction of ApoE would likely be detrimental to brain function. These results provide important insights into brain ApoE adaptive responses to increased lipid loads.

Dry eye disease (DED) is a chronic multifactorial disease of the ocular surface that can have a negative impact on quality of life and daily activities. This study aimed to elicit utility values for seven treatment health states of DED developed for use in an economic model to evaluate the benefits of DED treatment. DED health states were developed through interviews with clinicians (n=3) and patients (n=5), and tested through pilot time trade-off (TTO) exercises with members of the general population (n=5). Finalized DED health states were used to elicit utilities through vignette-based TTO interviews conducted in person with a sample from the UK general population (n=100). Seven hypothetical health states described different DED scenarios including descriptions of the symptoms, impacts, and treatment. These included health state A:"Initial Treatment Short Term", B:"Maintenance Strong Responder", C:"Maintenance Moderate Responder", D:"Long-term Post-initial Treatment", E:"Non-Responder", F:"Subsequent Treatments" and G:"Long-term Post-subsequent Treatment". Participants' ranked health states from most to least preferable, and then completed the TTO valuation exercise. Mean age of participants (n=100) was 38.8 years; 59% were female. Mean (standard deviation) EQ-5D-5L Visual Analogue Scale (VAS) and index scores were 85.1 (13.1) and 0.92 (0.12), respectively indicating a healthy sample. Health state B had the highest utility score (0.900) and was most commonly ranked the most preferable state followed by health state D (0.893). Health state G had the lowest score (0.656) and was most commonly ranked the least preferable health state. There were no significant differences in utility scores by age and gender between health states. Minor differences were seen between occupational groups. The findings suggest that responder health states are associated with markedly higher utility values. This evidence gives insight into the benefit of treatment for DED. Utility values elicited can be used in cost-effectiveness models comparing responses of treatment for DED.

This study was conducted to elicit utility values for the treatment of NMIBC, with the aim to understand preferences for different stages of bladder cancer for use in a cost-effectiveness model. Phase I developed and tested NMIBC health states, involving interviews with three clinicians, a patient advisor, and 30 members of the general population (as a pilot stage of the time trade-off [TTO] exercise). Interview findings were used to finalise the health states. Phase II involved utility elicitation; vignette-based TTO utility interviews conducted in-person with a sample of participants from the general population in London, United Kingdom (UK). Four chronic and one path hypothetical health states described different NMIBC scenarios. Participants were asked to rank each of the four chronic health states, followed by the TTO valuation exercise for all health states. Descriptions differed in terms of NMIBC symptoms’ severity, impacts, and mode and frequency of administration. Mean age of participants (n=202) was 46; 46% were female. Mean (standard deviation [SD]) EQ-5D-5L Visual Analogue Scale (VAS) and index scores were 83.2 (12.3) and 0.89 (0.18), respectively, indicating a healthy sample. Mean utilities were 0.781 for “No high-grade recurrence”, 0.586 for “High-grade recurrence”, 0.572 for “>1-year post cystectomy”, and 0.283 for “MIBC with metastatic disease”. The path health state of “First year post cystectomy” had a mean utility of 0.288. Pairwise comparisons found statistically significant differences between utilities (p < 0.001), except the comparison between high-grade recurrence and >1-year post cystectomy (p = 0.524). There were significant differences in health state utility scores by age and employment status. These results demonstrate that the general public highly value delaying radical cystectomy as a treatment alternative for bladder cancer. The health state with bladder-sparing therapy elicited a distinctly higher value compared to the other health states.

Brain health requires circuits, cells and molecular pathways to adapt when challenged and to promptly reset once the challenge has resolved. Neurodegeneration occurs when adaptability becomes confined, causing challenges to overwhelm neural circuitry. Studies of rare and common neurodegenerative diseases suggest that the accumulation of lipids can compromise circuit adaptability. Using microglia as an example, we review data that suggest increased lipid concentrations cause dysfunctional inflammatory responses to immune challenges, leading to Alzheimer's disease, Parkinson's disease and dementia. We highlight current approaches to treat lipid metabolic and clearance pathways and identify knowledge gaps towards restoring adaptive homeostasis in individuals who are at-risk of losing cognition.

Background: Lockdown and reconfiguration of NHS services imposed to limit the spread of SARS-CoV- 2 led to significantly reduced uptake of routine NHS care including access to cervical screening, contraceptive and sexual health services. We describe female sexual and reproductive health of a cohort of young women living with perinatally acquired HIV (YWLPaHIV) over the time-period covering SARS-CoV- 2 restrictions. Method: Single-centre observational cohort review of sexual and reproductive health for YWLPaHIV attending HIV services 01.07.2020 to 01.01.2021. Data collected included demographics, age of menarche/coitarche, Human papillomavirus (HPV) and Hepatitis B virus (HBV) vaccination, contraception, past sexually transmitted infections (STIs), cervical screening and obstetric history. Analysis was descriptive, with categorical analysis of qualitative data. Primary aim was to identify individuals in need of contraception, cervical screening and HBV vaccination/boosting. Results: 71 YWLPaHIV median age 23 years (IQR 21-27), 87% of black ethnicity, average body mass index 25.9kg/m2 (SD 5.6) with 58/71 (82%) latest viral load 10mIU. Conclusion: In this small cohort cervical smear uptake was below the national average despite high levels of previous abnormal findings. Given the high rates of prior STIs and pregnancy terminations, addressing the contraceptive and sexual health needs including vaccination for young women, may require accessible face-2- face services despite the pandemic.

To develop a conceptual disease model of health-related quality of life (HRQoL) in deaf children between the ages of 8 to 12 using cochlear implants (CIs). A targeted literature review was carried out to review research conducted in deaf children with CIs detailing concepts relevant to their HRQoL. The literature review informed two concept elicitation studies investigating HRQoL in the patient population. The interview guides focused on researching HRQoL concepts relevant to the patient experience of being deaf with CIs. The first study involved qualitative interviews conducted over the telephone with 7 professionals that have direct experience in working with the population which included: 2 clinical audiologists, 2 clinical psychologists, 1 speech therapist, 1 consultant surgeon and 1 principal teacher of the deaf. The second study involved face to face qualitative interviews with 15 deaf children with CIs between the ages of 8 and 12 years old. All interviews were conducted and transcribed verbatim and coded using thematic analysis being aided by QSR Nvivo 10 software. The analysis used an inductive bottom-up approach with the data driving analysis. The concepts researched in the literature review and the candidate themes found in analysis from the two studies were used to develop conceptual disease model domains that comprise HRQoL in deaf children with CIs. The interviews showed domains reported by both a wide range of professional and the children themselves. The final domains include: emotions, social relationships, school life, hearing functioning, and device limitations. Domains that were shown to be candidate themes but were not relevant included impact of family dynamic and identity of being a CI user. The conceptual HRQoL model presented has the potential to be used for development of a patient reported outcome measure assessing HRQoL in deaf children with CIs.

Cell and gene therapies (CGTs) are novel technologies poised to transform oncology treatment. In 2017, the U.S. Food and Drug Administration approved two CGTs for hematologic cancers. Social media provides a unique opportunity for patient-centric research to uncover what patients and clinicians value as important in their early experiences with CGTs. This study aimed to describe patient and clinician perceptions of CGT using publicly available social media forums in the U.S. Patient data included posts from oncology-specific patient forums and clinician data included transcripts from interviews with clinicians from open online sources. Data were extracted and anonymised, and further analysed using qualitative methods (thematic and content analyses). Emerging topics were coded and organised into themes and sub-themes. 302 patient posts across 6 patient forums dated between 2014-2017, and 35 clinician transcripts across 2 sites (OncoLive TV, Patient Power) between 2012- 2017 were included. The patient data review highlighted that patients often discussed CGT trial experiences and perceptions, CGT efficacy/safety, and shared scientific research findings from the literature and conferences. Forum users discussed challenges on entering a trial, as well as relayed CGT success stories. Long-term survival, efficacy, and side effects of CGT were discussed often in making decisions about enrolling in clinical trials. Reviewed clinician data included: experiences of trial efficacy/safety, concerns about increased costs (due to development, administration of CGTs, safety management), criteria for patient eligibility (age, fitness, cancer stage), and comparisons of CGTs with other immunotherapies and stem-cell transplants. Online forums are a vehicle for patients and physicians to express early experiences with CGT. Our analyses suggest that both groups are seeking information around several aspects of CGT and may benefit from increased education in these areas. Also, factors driving CGT treatment decisions should be further explored.

Autologous transplantation of patient-specific induced pluripotent stem cell (iPSC)-derived neurons is a potential clinical approach for treatment of neurological disease. Preclinical demonstration of long-term efficacy, feasibility, and safety of iPSC-derived dopamine neurons in non-human primate models will be an important step in clinical development of cell therapy. Here, we analyzed cynomolgus monkey (CM) iPSC-derived midbrain dopamine neurons for up to 2 years following autologous transplantation in a Parkinson’s disease (PD) model. In one animal, with the most successful protocol, we found that unilateral engraftment of CM-iPSCs could provide a gradual onset of functional motor improvement contralateral to the side of dopamine neuron transplantation, and increased motor activity, without a need for immunosuppression. Postmortem analyses demonstrated robust survival of midbrain-like dopaminergic neurons and extensive outgrowth into the transplanted putamen. Our proof of concept findings support further development of autologous iPSC-derived cell transplantation for treatment of PD.

To determine the long-term health and function of transplanted dopamine neurons in Parkinson’s disease (PD) patients, the expression of dopamine transporters (DATs) and mitochondrial morphology were examined in human fetal midbrain cellular transplants. DAT was robustly expressed in transplanted dopamine neuron terminals in the reinnervated host putamen and caudate for at least 14 years after transplantation. The transplanted dopamine neurons showed a healthy and nonatrophied morphology at all time points. Labeling of the mitochondrial outer membrane protein Tom20 and α-synuclein showed a typical cellular pathology in the patients’ own substantia nigra, which was not observed in transplanted dopamine neurons. These results show that the vast majority of transplanted neurons remain healthy for the long term in PD patients, consistent with clinical findings that fetal dopamine neuron transplants maintain function for up to 15–18 years in patients. These findings are critically important for the rational development of stem-cell-based dopamine neuronal replacement therapies for PD.

The main motor symptoms of Parkinson's disease are due to the loss of dopaminergic (DA) neurons in the ventral midbrain (VM). For the future treatment of Parkinson's disease with cell transplantation it is important to develop efficient differentiation methods for production of human iPSCs and hESCs-derived midbrain-type DA neurons. Here we describe an efficient differentiation and sorting strategy for DA neurons from both human ES/iPS cells and non-human primate iPSCs. The use of non-human primate iPSCs for neuronal differentiation and autologous transplantation is important for preclinical evaluation of safety and efficacy of stem cell-derived DA neurons. The aim of this study was to improve the safety of human- and non-human primate iPSC (PiPSC)-derived DA neurons. According to our results, NCAM(+) /CD29(low) sorting enriched VM DA neurons from pluripotent stem cell-derived neural cell populations. NCAM(+) /CD29(low) DA neurons were positive for FOXA2/TH and EN1/TH and this cell population had increased expression levels of FOXA2, LMX1A, TH, GIRK2, PITX3, EN1, NURR1 mRNA compared to unsorted neural cell populations. PiPSC-derived NCAM(+) /CD29(low) DA neurons were able to restore motor function of 6-hydroxydopamine (6-OHDA) lesioned rats 16 weeks after transplantation. The transplanted sorted cells also integrated in the rodent brain tissue, with robust TH+/hNCAM+ neuritic innervation of the host striatum. One year after autologous transplantation, the primate iPSC-derived neural cells survived in the striatum of one primate without any immunosuppression. These neural cell grafts contained FOXA2/TH-positive neurons in the graft site. This is an important proof of concept for the feasibility and safety of iPSC-derived cell transplantation therapies in the future.

Parkinson's disease associated mutations in leucine rich repeat kinase 2 (LRRK2) impair mitochondrial function and increase the vulnerability of induced pluripotent stem cell (iPSC)-derived neural cells from patients to oxidative stress. Since mitochondrial DNA (mtDNA) damage can compromise mitochondrial function, we examined whether LRRK2 mutations can induce damage to the mitochondrial genome. We found greater levels of mtDNA damage in iPSC-derived neural cells from patients carrying homozygous or heterozygous LRRK2 G2019S mutations, or at-risk individuals carrying the heterozygous LRRK2 R1441C mutation, than in cells from unrelated healthy subjects who do not carry LRRK2 mutations. After zinc finger nuclease-mediated repair of the LRRK2 G2019S mutation in iPSCs, mtDNA damage was no longer detected in differentiated neuroprogenitor and neural cells. Our results unambiguously link LRRK2 mutations to mtDNA damage and validate a new cellular phenotype that can be used for examining pathogenic mechanisms and screening therapeutic strategies.

Alternative splicing is a complex post-transcriptional process that can be regulated by cis-acting elements located within genomic non-coding regions. Recent studies have identified that polymorphic variations in non-coding regions of the α-synuclein gene (SNCA) locus are associated with an increased risk for developing Parkinson's disease (PD). The underlying mechanism(s) for this susceptibility may involve changes in α-synuclein mRNA expression and alternative splicing. As a first step towards understanding the biology of α-synuclein splice variants in PD, we characterized the levels of the full-length SNCA-140 mRNA transcript and SNCA-126, -112, and -98 alternatively spliced variants in different neuronal regions from PD patients or transgenic mice overexpressing human α-synuclein (ASO). In human post-mortem tissue, α-synuclein spliced transcripts were expressed in a region-specific manner in the cortex, substantia nigra, and cerebellum. We observed increased nigral SNCA-140 and SNCA-126 transcript levels in PD patients when compared to neurologically unaffected cases. Human α-synuclein splicing changes were also found to occur in a region-specific manner in ASO mice. Here, SNCA-126, -112, and -98 transcript levels did not increase proportionally with SNCA-140 levels, or parallel the region-specific mouse transcript ratios seen in wild-type (WT) littermates. While most transcripts were elevated in ASO mice when compared to WT mice, the most prominent increase was found in the ventral midbrain of 15-month-old ASO mice. These results demonstrate region-specific human α-synuclein transcript level abnormalities in PD patients and in a transgenic mouse model of α-synucleinopathy. This study is relevant to understanding the normal, adaptive, or pathological role(s) of α-synuclein splice variants.