BACKGROUND. NAFLD affects 25-30% of the US and European populations and is associated with insulin resistance (IR), T2D, increased cardiovascular risk and is defined by hepatic triglyceride content (HTG) > 5.56%. However, it is unknown whether HTG content less than 5.56% is associated with cardiometabolic risk factors and whether there are ethnic [Asian Indian (AI) vs. non-Asian Indian (non-AI)] and/or gender differences in these parameters in lean individuals. METHODS. We prospectively recruited 2,331 individuals and measured HTG, using 1H MRS, and plasma concentrations of triglycerides, total cholesterol, LDL cholesterol, HDL cholesterol, and uric acid. Insulin sensitivity was assessed using HOMA-IR and the Matsuda Insulin Sensitivity Index (ISI). RESULTS. The 95th percentile for HTG in lean non-AI individuals was 1.85%. Plasma insulin, triglycerides, total cholesterol, LDL cholesterol and uric acid concentrations were increased and HDL decreased in individuals with HTG content > 1.85% and ≤ 5.56% compared to those individuals with HTG content ≤ 1.85% and was associated with increased IR. Mean HTG was lower in lean non-AI women compared to lean non-AI men, whereas lean AI men and women had a 40-100% increase in HTG when compared to non-AI men and women which was associated with increased cardiometabolic risk factors. CONCLUSIONS. We found that the 95th percentile of HTG in lean non-AI individuals was 1.85% and that HTG concentrations above this threshold were associated with IR and cardiovascular risk factors. Premenopausal women are protected from these changes whereas young lean AI men and women manifest increased HTG content and associated cardiometabolic risk factors. FUNDING. Supported by grants from the United States Department of Health and Human Resources (NIH/NIDDK): R01 DK113984, P30 DK45735, U24 DK59635 and UL1 RR024139 and the Novo Nordisk Foundation (NNF18CC0034900).
Kitt Falk Petersen, Sylvie Dufour, Fangyong Li, Douglas L. Rothman, Gerald I. Shulman
BACKGROUND. Pathophysiology of type 1 diabetes (T1D) is illustrated by pancreatic islet infiltration of inflammatory lymphocytes, including CD8+ T-cells; however, the molecular factors mediating their recruitment remain unknown. We hypothesized that single-cell RNA-sequencing (scRNA-Seq) analysis of immune cell populations isolated from islets of non-obese diabetic (NOD) mice captured gene expression dynamics providing critical insight into autoimmune diabetes pathogenesis. METHODS. Pancreatic sections from human donors were investigated, including T1D subjects, auto-antibody-positive (aAb+), and non-diabetic controls. Immunohistochemistry was performed to assess islet hormones, and both novel and canonical immune-cell markers that were identified from state-of-the-art workflows after reanalyzing murine scRNA-seq datasets. RESULTS. Computational workflows identified Cadm1-mediated binding among the most significant intercellular interactions among all cell clusters as well as Cadm1 enrichment in macrophages and dendritic cells from pancreata of NOD mice. Immunostaining of human pancreata revealed an increased number of CADM1+GCG+ cells adjacent to CD8+ T-cells in sections from T1D and aAb+ donors compared to non-diabetic subjects. Numbers of CADM1+CD68+ peri-islet myeloid cells adjacent to CD8+ T-cells were also increased in pancreatic sections from both T1D and aAb+ donors compared to non-diabetic subjects. CONCLUSION. Increased detection of CADM1+ cells adjacent to CD8+ T-cells in pancreatic sections of T1D and aAb+ human subjects validated workflows, and indicates CADM1-mediated intercellular contact may facilitate islet infiltration of cytotoxic T lymphocytes and serve as a potential therapeutic target for preventing T1D pathogenesis. FUNDING. The Johns Hopkins All Children’s Foundation IRG Program, NSFC (82071326) and DFG (431549029–SFB 1451, EXC 2030–390661388, and 411422114-GRK 2550).
Chandan Sona, Yu-Te Yeh, Andreas Patsalos, Laszlo Halasz, Xin Yan, Natalia L. Kononenko, Laszlo Nagy, Matthew N. Poy
BACKGROUND. Paclitaxel chemotherapy frequently induces dose-limiting sensory axonal polyneuropathy. As sensory symptoms are challenging to assess objectively in clinical routine, an easily accessible biomarker for chemotherapy-induced polyneuropathy (CIPN) holds the potential to improve early diagnosis. Here, we describe neurofilament light chain (NFL), a marker for neuroaxonal damage, as translational surrogate marker for CIPN. METHODS. NFL concentrations were measured in an in vitro model of CIPN, exposing induced pluripotent stem cell-derived sensory neurons (iPSC-DSN) to paclitaxel. Breast and ovarian cancer patients undergoing paclitaxel chemotherapy, breast cancer control patients without chemotherapy and healthy controls were recruited in a cohort study and examined before chemotherapy (V1) and after 28 weeks (V2, after chemotherapy). CIPN was assessed by the validated Total Neuropathy Score reduced, which combines patient-reported symptoms with data from clinical examinations. Serum NFL (NFLs) concentrations were measured at both visits with single molecule array technology (SIMOA). RESULTS. NFL is released from iPSC-DSN upon paclitaxel incubation in a dose- and time-dependent manner and inversely correlates with iPSC-DSN viability. NFLs strongly increased in paclitaxel-treated patients with CIPN, but not in chemotherapy patients without CIPN or controls, resulting in an 86 % sensitivity and 87 % specificity. A NFLs increase of +36 pg/ml from baseline was associated with a predicted CIPN probability of >0.5. CONCLUSION. NFLs correlates with CIPN development and severity, which may guide neurotoxic chemotherapy in the future. TRIAL REGISTRATION. NCT02753036 FUNDING. DFG (EXC 257 NeuroCure), BMBF (01 EO 0801), AnimalFreeResearch Organization, EU Horizon 2020 Innovative Medicines Initiative 2 Joint Undertaking (TransBioLine, 821283)
Petra Huehnchen, Christian Schinke, Nikola Bangemann, Adam D. Dordevic, Johannes Kern, Smilla K. Maierhof, Lois Hew, Luca Nolte, Peter Körtvelyessy, Jens C. Göpfert, Klemens Ruprecht, Christopher J. Somps, Jens-Uwe Blohmer, Jalid Sehouli, Matthias Endres, Wolfgang Boehmerle
BACKGROUND. Accumulation of advanced glycation endproducts (AGEs) may contribute to the pathophysiology of type 2 diabetes and its vascular complications. AGEs are widely present in food, but whether restricting AGE intake improves risk factors for type 2 diabetes and vascular dysfunction is controversial. RESEARCH DESIGN AND METHODS. Abdominally obese but otherwise healthy individuals were randomly assigned to a specifically designed 4-week diet low or high in AGEs in a double blind parallel-design. Insulin sensitivity, secretion, and clearance were assessed by a combined hyperinsulinemic-euglycemic and hyperglycemic clamp. Micro- and macrovascular function, inflammation, and lipid profile were assessed by state-of-art in vivo measurements and biomarkers. Specific urinary and plasma AGEs Nε-(carboxymethyl)lysine (CML), Nε-(1-carboxyethyl)lysine (CEL), and Nδ-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine (MG-H1) were assessed by mass spectrometry. RESULTS. In 73 individuals (22 males, mean ± SD age and BMI 52 y ± 14, 30.6 kg/m2 ± 4.0), intake of CML, CEL, and MG-H1 differed 2.7, 5.3, and 3.7-fold between the low and high AGE diets, which led to corresponding changes of these AGEs in urine and plasma. Despite this, there was no difference in insulin sensitivity, secretion, or clearance, micro- and macrovascular function, overall inflammation, or lipid profile between the low and high dietary AGE groups (all p for treatment effects > 0.05). CONCLUSIONS. This comprehensive RCT demonstrates very limited biological consequences of a 4-week diet low or high in AGEs in abdominally obese individuals. TRIAL REGISTRATION. clinicaltrials.gov: NCT03866343, trialregister.nl: NTR7594. FUNDING. Diabetesfonds and ZonMw.
Armand M A Linkens, Alfons J. Houben, Petra M Niessen, Nicole Wijckmans, Erica de Goei, Mathias D.G. Van den Eynde, Jean L. J. M. Scheijen, Marjo Waarenburg, Andrea Mari, Tos T.J.M. Berendschot, Lukas Streese, Henner Hanssen, Martien C.J.M. van Dongen, Christel van Gool, Coen D.A. Stehouwer, Simone JPM Eussen, Casper Schalkwijk.
BACKGROUND Outcome measures sensitive to disease progression are needed for ATP-binding cassette, sub-family A, member 4–associated (ABCA4-associated) retinopathy. We aimed to quantify ellipsoid zone (EZ) loss and photoreceptor degeneration beyond EZ-loss in ABCA4-associated retinopathy and investigate associations between photoreceptor degeneration, genotype, and age.METHODS We analyzed 132 eyes from 66 patients (of 67 enrolled) with molecularly confirmed ABCA4-associated retinopathy from a prospective natural history study with a median [IQR] follow-up of 4.2 years [3.1, 5.1]. Longitudinal spectral-domain optical coherence tomography volume scans (37 B-scans, 30° × 15°) were segmented using a deep learning (DL) approach. For genotype-phenotype analysis, a model of ABCA4 variants was applied with the age of criterion EZ-loss (6.25 mm2) as the dependent variable.RESULTS Patients exhibited an average (square-root-transformed) EZ-loss progression rate of [95% CI] 0.09 mm/y [0.06, 0.11]. Outer nuclear layer (ONL) thinning extended beyond the area of EZ-loss. The average distance from the EZ-loss boundary to normalization of ONL thickness (to ±2 z score units) was 3.20° [2.53, 3.87]. Inner segment (IS) and outer segment (OS) thinning was less pronounced, with an average distance from the EZ-loss boundary to layer thickness normalization of 1.20° [0.91, 1.48] for the IS and 0.60° [0.49, 0.72] for the OS. An additive model of allele severity explained 52.7% of variability in the age of criterion EZ-loss.CONCLUSION Patients with ABCA4-associated retinopathy exhibited significant alterations of photoreceptors outside of EZ-loss. DL-based analysis of photoreceptor laminae may help monitor disease progression and estimate the severity of ABCA4 variants.TRIAL REGISTRATION ClinicalTrials.gov identifier: NCT01736293.FUNDING National Eye Institute Intramural Research Program and German Research Foundation grant PF950/1-1.
Maximilian Pfau, Catherine A. Cukras, Laryssa A. Huryn, Wadih M. Zein, Ehsan Ullah, Marisa P. Boyle, Amy Turriff, Michelle A. Chen, Aarti S. Hinduja, Hermann E.A. Siebel, Robert B. Hufnagel, Brett G. Jeffrey, Brian P. Brooks
BACKGROUND. Prostate cancer is multifocal with distinct molecular subtypes. The utility of genomic subtyping has been challenged due to inter- and intra-focal heterogeneity. We sought to characterize the subtype-defining molecular alterations of primary prostate cancer across all tumor foci within radical prostatectomy (RP) specimens and determine the prevalence of collision tumors. METHODS. From the Early Detection Research Network cohort, we identified 333 prospectively collected RPs from 2010 to 2014 and assessed ERG, SPINK1, PTEN, and SPOP molecular status. We utilized dual ERG/SPINK1 immunohistochemistry, fluorescence in situ hybridization to confirm ERG rearrangements and characterize PTEN deletion, and high-resolution melting curve analysis and Sanger sequencing to determine SPOP mutation status. Analysis of biochemical recurrence-free of patients with collision tumors was conducted using Kaplan-Meier method. RESULTS. Based on index focus alone, ERG, SPINK1, PTEN, and SPOP alterations were identified in 47.5%, 10.8%, 14.3%, and 5.1% of RP specimens, respectively. In 233 multifocal RPs with ERG/SPINK1 status in all foci, 139 (59.7%) had discordant molecular alterations between foci. Collision tumors, as defined by discrepant ERG/SPINK1 status within a single focus, were identified in 29 (9.4%) RP specimens. CONCLUSION. Interfocal molecular heterogeneity was identified in ~60% of multifocal RP specimens and collision tumors were present in ~10%. We present this phenomenon as a model for the intra-focal heterogeneity observed in previous studies and propose future genomic studies screen for collision tumors to better characterize molecular heterogeneity.
Jacqueline Fontugne, Peter Y. Cai, Hussein Alnajar, Bhavneet Bhinder, Kyung Park, Huihui Ye, Shaham Beg, Verena Sailer, Javed Siddiqui, Mirjam Blattner-Johnson, Jaclyn A. Croyle, Zohal Noorzad, Carla Calagua, Theresa Y. MacDonald, Ulrika Axcrona, Mari Bogaard, Karol Axcrona, Douglas S. Scherr, Martin G. Sanda, Bjarne Johannessen, Arul M. Chinnaiyan, Olivier Elemento, Rolf I. Skotheim, Mark A. Rubin, Christopher E. Barbieri, Juan M. Mosquera
BACKGROUND. Vaccine-elicited adaptive immunity is a prerequisite for control of SARS-CoV-2 infection. Multiple sclerosis (MS) disease-modifying therapies (DMTs) differentially target humoral and cellular immunity. A comprehensive comparison of MS DMTs on SARS-CoV-2 vaccine-specific immunity is needed, including quantitative and functional B and T cell responses. METHODS. Spike-specific antibody and T cell responses were measured before and following SARS-CoV-2 vaccination in a cohort of 80 subjects, including healthy controls and MS patients in six DMT groups: untreated, glatiramer acetate (GA), dimethyl fumarate (DMF), natalizumab (NTZ), sphingosine-1-phosphate (S1P) receptor modulators, and anti-CD20 monoclonal antibodies. Anti-spike antibody responses were quantified by Luminex assay, high-resolution spike epitope reactivity was mapped by VirScan, and pseudovirus neutralization was assessed. Spike-specific CD4+ and CD8+ T cell responses were characterized by activation-induced marker (AIM) expression, cytokine production, and tetramer analysis. RESULTS. Anti-spike IgG levels were similar between healthy controls, untreated MS, GA, DMF, and NTZ patients, but were significantly reduced in anti-CD20 and S1P-treated patients. Anti-spike seropositivity in anti-CD20 patients was significantly correlated with CD19+ B cell levels and inversely correlated with cumulative treatment duration. Spike epitope reactivity and pseudovirus neutralization was reduced in anti-CD20 and S1P patients, directly correlating with reduced spike receptor binding domain (RBD) IgG levels. Spike-specific CD4+ and CD8+ T cell reactivity remained robust across all groups except in S1P-treated patients in whom post-vaccine CD4+ T cell responses were attenuated. CONCLUSIONS. These findings from a large MS cohort exposed to a wide spectrum of MS immunotherapies have important implications for treatment-specific COVID-19 clinical guidelines. FUNDING. This work was supported by grants from the NIH 1K08NS107619 (JJS), NMSS TA- 1903-33713 (JJS), K08NS096117 (MRW), Westridge Foundation (MRW), Chan Zuckerberg Biohub (JLD), R01AI159260 (JAH), R01NS092835 (SSZ), R01AI131624 (SSZ), R21NS108159 (SSZ), NMSS RG1701-26628 (SSZ), and the Maisin Foundation (SSZ).
Joseph J. Sabatino Jr, Kristen Mittl, William M. Rowles, Kira McPolin, Jayant V. Rajan, Matthew T. Laurie, Colin R. Zamecnik, Ravi Dandekar, Bonny D. Alvarenga, Rita P. Loudermilk, Chloe Gerungan, Collin M. Spencer, Sharon A. Sagan, Danillo G. Augusto, Jessa R. Alexander, Joseph L. DeRisi, Jill A. Hollenbach, Michael R. Wilson, Scott S. Zamvil, Riley Bove
BACKGROUND. Most subjects with prior COVID-19 disease manifest long–term, protective immune responses against re-infection. Accordingly, we tested the hypothesis that humoral immune and reactogenicity responses to a SARS-CoV-2 mRNA vaccine differ in subjects with and without prior COVID-19. METHODS. Health care workers (n=61) with (n=30) and without (n=31) prior COVID-19 disease received two, 30 µg doses of Pfizer BNT162b2 vaccine 3 weeks apart. Serum IgG antibody against the Spike receptor-binding domain (RBD); serum neutralizing activity; and vaccine reactogenicity were assessed longitudinally every 2 weeks for 56 days after the 1st injection. RESULTS. The COVID group manifested more rapid increases in Spike IgG antibody and serum neutralizing activity post 1st vaccine dose but showed little or no increase after the 2nd dose compared to the infection-naïve group. In fact, Spike IgG was maximum after the 1st dose in 36% of the COVID group versus 0% of the infection-naïve group. Peak IgG antibody was lower but appeared to fall more slowly in the COVID-19 versus the infection-naïve group. Finally, adverse systemic reactions e.g., fever, headache and malaise, were more frequent and lasted longer after both the 1st and 2nd injection in the COVID group than in the infection-naïve group. CONCLUSION. Subjects with prior COVID-19 demonstrate a robust, accelerated humoral immune response to the 1st dose but attenuated response to the 2nd dose of BNT162b2 vaccine compared to controls. The COVID-19 group also experiences greater reactogenicity. Humoral responses and reactogenicity to BNT162b2 differ qualitatively and quantitatively in subjects with prior COVID-19 compared to infection-naive subjects. FUNDING. This work was supported by Institutional Funds.
Steven G. Kelsen, Alan S. Braverman, Mark O. Aksoy, Jacob A. Hayman, Puja S. Patel, Charu Rajput, Huaqing Zhao, Susan G. Fisher, Michael R. Ruggieri Sr., Nina T. Gentile
BACKGROUND. Adenoviral (Ad)-vectored vaccines are typically administered via intramuscular injection to humans, incapable of inducing respiratory mucosal immunity. However, aerosol delivery of Ad-vectored vaccines remains poorly characterized and its ability to induce mucosal immunity in humans is unknown. This phase 1b trial was to evaluate the safety and immunogenicity of human serotype-5 Ad-vectored tuberculosis (TB) vaccine (AdHu5Ag85A) delivered to humans via inhaled aerosol or intramuscular injection. METHODS. 31 healthy, previously BCG-vaccinated adults were enrolled. AdHu5Ag85A was administered by single-dose aerosol using Aeroneb® Solo Nebulizer or by intramuscular (IM) injection. The study consisted of the low dose (LD) aerosol, high dose (HD) aerosol and IM groups. The adverse events were assessed at various times post-vaccination. Immunogenicity data were collected from the peripheral blood and bronchoalveolar lavage samples at baseline and select timepoints post-vaccination. RESULTS. The nebulized aerosol droplets were <5.39µm in size. Both LD and HD of AdHu5Ag85A administered by aerosol inhalation and IM injection were safe and well-tolerated. Both aerosol doses, particularly LD, but not IM, vaccination markedly induced airway tissue-resident memory CD4 and CD8 T cells of polyfunctionality. While as expected, IM vaccination induced Ag85A-specific T cell responses in the blood, the LD aerosol vaccination also elicited such T cells in the blood. Furthermore, the LD aerosol vaccination induced persisting transcriptional changes in alveolar macrophages. CONCLUSIONS. Inhaled aerosol delivery of Ad-vectored vaccine is a safe and superior way to elicit respiratory mucosal immunity. This study warrants further development of aerosol vaccine strategies against respiratory pathogens including TB and COVID-19. TRIAL REGISTRATION. This trial is registered with ClinicalTrial.gov, NCT# 02337270. FUNDING. The Canadian Institutes for Health Research and the Natural Sciences and Engineering Research Council of Canada.
Mangalakumari Jeyanathan, Dominik K. Fritz, Sam Afkhami, Emilio Aguirre, Karen J. Howie, Anna Zganiacz, Anna Dvorkin-Gheva, Michael R. Thompson, Richard Silver, Ruth P. Cusack, Brian D. Lichty, Paul M. O'Byrne, Martin Kolb, Maria Fe C. Medina, Myrna B. Dolovich, Imran Satia, Gail M Gauvreau, Zhou Xing, Fiona Smaill
BACKGROUND. >1,500 variants in the ABCA4 locus underlie a heterogeneous spectrum of retinal disorders ranging from aggressive childhood-onset chorioretinopathy to milder, late-onset macular disease. Genotype-phenotype correlation studies have been limited in clinical applicability as patient cohorts are typically small and seldom capture the full natural history of individual genotypes. To overcome these limitations, we constructed a genotype-phenotype correlation matrix that provides quantifiable probabilities of long-term disease outcomes associated with specific ABCA4 genotypes from a large, age-restricted patient cohort. METHODS. The study included 112 unrelated patients ≥50 years of age in whom 2 pathogenic variants were identified after sequencing of the ABCA4 locus. Clinical characterization was performed using the results of best-corrected visual acuity, retinal imaging and full-field electroretinogram testing. RESULTS. Four distinct prognostic groups were defined according to the spatial severity of disease features across the fundus. Recurring genotypes were observed in milder prognoses including those associated with a newly defined class of rare hypomorphic alleles. PVS1 (predicted null) variants were enriched in the most severe prognoses; however, missense variants comprised a larger than expected fraction of these patients. Analysis of allele combinations and their respective prognostic severity, showed that certain variants such as p.(Gly1961Glu), and both rare and frequent hypomorphic alleles, are “clinically dominant” with respect to patient phenotypes irrespective of the allele in trans. CONCLUSION. These results provide much needed structure to the complex genetic and clinical landscape of ABCA4 disease and adds a tool to the clinical repertoire to quantitatively assess individual genotype-specific prognoses in patients.
Winston Lee, Jana Zernant, Pei-Yin Su, Takayuki Nagasaki, Stephen H. Tsang, Rando Allikmets
No posts were found with this tag.