Data_Sheet_1_Mendelian randomization analysis suggests no associations of human herpes viruses with amyotrophic lateral sclerosis.doc

BackgroundThe causal associations between infections with human herpes viruses (HHVs) and amyotrophic lateral sclerosis (ALS) has been disputed. This study investigated the causal associations between herpes simplex virus (HSV), varicella-zoster virus (VZV), Epstein–Barr virus (EBV), cytomegalovirus (CMV), HHV-6, and HHV-7 infections and ALS through a bidirectional Mendelian randomization (MR) method.MethodsThe genome-wide association studies (GWAS) database were analyzed by inverse variance weighted (IVW), MR-Egger, weighted median, simple mode, and weighted mode methods. MR-Egger intercept test, MR-PRESSO test, Cochran’s Q test, funnel plots, and leaveone-out analysis were used to verify the validity and robustness of the MR results.ResultsIn the forward MR analysis of the IVW, genetically predicted HSV infections [odds ratio (OR) = 0.9917; 95% confidence interval (CI): 0.9685–1.0154; p = 0.4886], HSV keratitis and keratoconjunctivitis (OR = 0.9897; 95% CI: 0.9739–1.0059; p = 0.2107), anogenital HSV infection (OR = 1.0062; 95% CI: 0.9826–1.0304; p = 0.6081), VZV IgG (OR = 1.0003; 95% CI: 0.9849–1.0160; p = 0.9659), EBV IgG (OR = 0.9509; 95% CI: 0.8879–1.0183; p = 0.1497), CMV (OR = 0.9481; 95% CI: 0.8680–1.0357; p = 0.2374), HHV-6 IgG (OR = 0.9884; 95% CI: 0.9486–1.0298; p = 0.5765) and HHV-7 IgG (OR = 0.9991; 95% CI: 0.9693–1.0299; p = 0.9557) were not causally associated with ALS. The reverse MR analysis of the IVW revealed comparable findings, indicating no link between HHVs infections and ALS. The reliability and validity of the findings were verified by the sensitivity analysis.ConclusionAccording to the MR study, there is no evidence of causal associations between genetically predicted HHVs (HSV, VZV, EBV, CMV, HHV-6, and HHV-7) and ALS.</p

Phenotype of TPBG Gene Replacement in the Mouse and Impact on the Pharmacokinetics of an Antibody–Drug Conjugate

The use of predictive preclinical models in drug discovery is critical for compound selection, optimization, preclinical to clinical translation, and strategic decision-making. Trophoblast glycoprotein (TPBG), also known as 5T4, is the therapeutic target of several anticancer agents currently in clinical development, largely due to its high expression in tumors and low expression in normal adult tissues. In this study, mice were engineered to express human TPBG under endogenous regulatory sequences by replacement of the murine Tpbg coding sequence. The gene replacement was considered functional since the hTPBG knockin (hTPBG-KI) mice did not exhibit clinical observations or histopathological phenotypes that are associated with Tpbg gene deletion, except in rare instances. The expression of hTPBG in certain epithelial cell types and in different microregions of the brain and spinal cord was consistent with previously reported phenotypes and expression patterns. In pharmacokinetic studies, the exposure of a clinical-stage anti-TPBG antibody–drug conjugate (ADC), A1mcMMAF, was lower in hTPBG-KI versus wild-type animals, which was evidence of target-related increased clearance in hTPBG-KI mice. Thus, the hTPBG-KI mice constitute an improved system for pharmacology studies with current and future TPBG-targeted therapies and can generate more precise pharmacokinetic and pharmacodynamic data. In general the strategy of employing gene replacement to improve pharmacokinetic assessments should be broadly applicable to the discovery and development of ADCs and other biotherapeutics

Long-chain solid organic polysulfide cathode for high-capacity secondary lithium batteries

Organic polysulfides are linear sulfur chains (R–S–R, n ≥ 2) capped with organic moieties, and are appealing cathode materials in lithium batteries. The theoretical capacity of polysulfides essentially relies on the length of the sulfur chains; long-chain polysulfides could store more charges than short-chain polysulfides. Herein, we report the successful synthesis of a long-chain solid organic polysulfide (SOPS) by a radical coupling method and disclose its functions as high capacity cathode materials for secondary lithium batteries. The capped, long-chain polysulfide offers the fast charge transfer and thereby enabling the better performance than the pure sulfur. The SOPS cathode reaches an initial capacity of 1166 mA h g at 750 mA g, and maintains a high capacity of 674 mA h g at 3000 mA g

Serine Protease HTRA1 Antagonizes Transforming Growth Factor-β Signaling by Cleaving Its Receptors and Loss of HTRA1 <i>In Vivo</i> Enhances Bone Formation

<div><p>HTRA1 is a member of the High Temperature Requirement (HTRA1) family of serine proteases, which play a role in several biological and pathological processes. In part, HTRA1 regulation occurs by inhibiting the TGF-β signaling pathway, however the mechanism of inhibition has not been fully defined. Previous studies have shown that HTRA1 is expressed in a variety of tissues, including sites of skeletal development. HTRA1 has also been implicated in the process of bone formation, although the precise manner of regulation is still unknown. This study investigated how HTRA1 regulates TGF-β signaling and examined the <i>in vivo</i> effects of the loss of HTRA1. We demonstrated that recombinant HTRA1 was capable of cleaving both type II and type III TGF-β receptors (TβRII and TβRIII) <i>in vitro</i> in a dose-dependent manner, but it did not affect the integrity of TβRI or TGF-β. Overexpression of HTRA1 led to decreased levels of both TβRII and III on the cell surface but had no effect on TβRI. Silencing HTRA1 expression significantly increased TGF-β binding to the cell surface and TGF-β responsiveness within the cell. To examine the role of HTRA1 <i>in vivo</i>, we generated mice with a targeted gene deletion of <i>HTRA1</i>. Embryonic fibroblasts isolated from these mice displayed an increase in TGF-β-induced expression of several genes known to promote bone formation. Importantly, the loss of HTRA1 in the knockout mice resulted in a marked increase in trabecular bone mass. This study has identified a novel regulatory mechanism by which HTRA1 antagonizes TGF-β signaling, and has shown that HTRA1 plays a key role in the regulation of bone formation.</p></div

Regulation of TGF-β signaling by HTRA1.

<p>(<i>A</i>)(<i>B</i>) Binding of TGF-β to the cell surface as measured by flow cytometry. Cells were transfected with either nonspecific control siRNA or HTRA1 siRNA and then incubated with either biotinylated TGF-β or a biotinylated negative control protein to measure the background. Solid gray = background, dashed line = control siRNA, black line = HTRA1 siRNA. (<i>B</i>) Bar chart indicates the mean FITC values after subtraction of the background level. **<i>P</i><0.01, Student’s <i>t</i>-test, n = 4. (<i>C</i>) Immunoblot of total and phosphorylated Smad2 (465/467) in A549 cells transfected with either control or HTRA1 siRNA, and then treated with 1 ng/ml TGF-β for the indicated times. β-actin was used as a loading control. (<i>D</i>) The effect of HTRA1 knockdown on the expression of TGF-β-regulated genes. A549 cells were transfected with either nonspecific control siRNA or HTRA1 siRNA and then treated with 1 ng/ml TGF-β for the indicated times. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, Student’s <i>t</i>-test, n = 3.</p

The <i>in vivo</i> effects of loss of HTRA1.

<p>(<i>A</i>) Schematic of the <i>HTRA1</i> knockout strategy. Primers were designed with one forward primer (For) that amplifies an amplicon for the wild type and heterozygous alleles with a corresponding reverse primer (Rev), and a second reverse primer designed to only amplify the knockout. The knockout reverse primer (KO Rev) only amplifies following CRE-LOX recombination. Solid triangles = loxP sites; EV = EcoRI. E2 = Exon2; E3 = Exon3. (<i>B</i>) Genotype results of mouse embryonic fibroblasts showing confirmation of the generation of <i>HTRA1</i>+/− and <i>HTRA1</i>−/− mice. (<i>C</i>) Expression of TGF-β-regulated genes in embryonic fibroblasts from wild type (WT) and <i>HTRA1</i> knockout (KO) mice. Cells were treated with 1 ng/ml TGF-β for the indicated times. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, Student’s <i>t</i>-test, n = 9 for each cell type. (<i>D</i>) Microcomputed tomography and parameters of trabecular bone in the distal femurs in wild type (WT), <i>HTRA1</i> heterozygous (HET) and <i>HTRA1</i> knockout (KO) mice (all males, 3 months old). (<i>E</i>) Microcomputed tomography and parameters of trabecular bone in the fourth vertebrae in wild type (WT), <i>HTRA1</i> heterozygous (HET) and <i>HTRA1</i> knockout (KO) mice. For both (<i>D</i>) and (<i>E</i>), *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, ANOVA and Fisher’s protected least significant difference test, n = 13.</p