SynZyme Technologies (United States)

Enzymes accelerate biochemical reactions in living organisms, thus playing an important role in metabolism. Although metabolic pathway databases are growing, many metabolic reactions, termed orphan enzymes, have not been annotated to gene sequences, which hinders functional annotation in genomic analysis. Moreover, protein databases contain many proteins of unknown function. Owing to this gap between known proteins and enzymatic reactions, various proteins of unknown function may be orphan enzymes; however, available tools cannot adequately predict these links. In this study, we developed DeepRES, an AI-based framework for comprehensive enzyme screening, to explore novel enzyme candidates from proteins of unknown function for reactions of interest. DeepRES implements enzyme screening via two steps: classification of enzymes and non-enzymes and prediction of catalytic capabilities for enzyme‒reaction pairs. The two deep learning models comprising DeepRES showed comparable or superior performance to that of existing software. We performed screening of 1,255 orphan enzymes involved in the microbiome using DeepRES and successfully identified candidate proteins for 897 orphan enzymes. We then used those candidates as references for genomic analysis and explored novel biosynthetic gene clusters from microbial genomes to obtain promising candidate gene clusters, including those related to anthocyanin degradation. Comprehensive enzyme screening via DeepRES, which is the first computational tool designed to associate orphan enzymes with proteins of unknown function, is expected to facilitate high-throughput identification of orphan enzyme-encoding genes. Furthermore, DeepRES can be easily integrated into the current genomic analysis pipeline to extend the functional annotation.

Abstract Introduction Oral testosterone undecanoate (TU) was approved outside the United States 50 years ago and is currently used in several countries for the treatment of men with testosterone deficiency. The efficacy of prior oral TU is limited by a short half-life, requiring frequent dosing and administration with meals. A novel formulation of fixed-dose TU is now indicated for the treatment of testosterone deficiency (FDA approved March 2022). In a phase 3 study (NCT03242590: 1021-16-002) evaluating the efficacy and safety of fixed-dose oral TU 225 mg twice daily in adult men with testosterone deficiency, oral TU was well tolerated and restored testosterone levels to eugonadal ranges in most patients. Objective The objective of this pooled analysis of ten clinical studies (LPCN 1021-09-001, S361.1.001, M12-778, M13-298, 1021-14-001, LPCN 1021-16-002, LPCN 1021-16-003, LPCN 1021-13-001, LPCN 1021-18-001, LPCN 1021-18-003) was to further evaluate the safety of fixed-dose TU administered to adult men with testosterone deficiency. Methods This pooled analysis included men with testosterone deficiency who were enrolled or randomized in one of ten clinical studies and received at least one dose of study drug, which included fixed-dose TU, dose-adjusted TU, or topical testosterone gel. Data from separate studies were allocated to defined treatment groups, and these data were pooled to present a safety summary of patients treated with fixed-dose TU. Treatment groups were defined as follows: all fixed-dose TU patients, active control patients (dose-adjusted TU or topical testosterone gel), and placebo patients. Participants in a crossover study of various fixed-dose TU doses were counted once overall for analysis, and data relating to the highest dose were used. Results Overall, 777 patients were included in this pooled analysis: 654 received fixed-dose TU, 104 received topical testosterone gel, 34 received dose-adjusted TU, and 18 received placebo. Baseline characteristics are available on Table 1. Among 654 patients who received fixed-dose TU, 549 (83.9%) patients completed their respective study, and 15 (2.3%) patients discontinued because of an AE. The most frequently occurring treatment-emergent related adverse events (AEs) for patients treated with fixed-dose TU included headache (10 [1.5]), hematocrit increase (8 [1.2]), and acne (6 [0.9]) (Table 2). Three (0.5%) patients receiving fixed-dose TU experienced a blood pressure increase and 13 (2.0%) exhibited hypertension. No patients receiving fixed-dose TU experienced an increase in hepatic enzymes. The mean change from baseline (SD) in hematocrit (HCT) for patients receiving fixed-dose TU was 0.003% (0.029); three patients receiving fixed-dose TU discontinued because of HCT >54%. No patients in this pooled analysis experienced a serious AE (SAE) related to study treatment, and no deaths occurred. Conclusions In this pooled analysis of ten clinical studies in male patients with testosterone deficiency, fixed-dose TU was well tolerated with no hepatic adverse events, no drug-related SAEs, and no deaths. Disclosure Yes, this is sponsored by industry/sponsor: Halozyme (previously Antares) Clarification Industry initiated, executed and funded study Any of the authors act as a consultant, employee or shareholder of an industry for: Halozyme (previously Antares)

BackgroundTroponin is an important marker for the diagnosis of acute myocardial infarction (AMI). The detection of troponin in peripheral blood is simpler and more convenient than that in venous blood, which has attracted more and more clinical attention. The purpose of this study is to establish a novel method for the rapid detection of high-sensitivity troponin I (hs-cTnI) in peripheral blood by quantum dot fluorescence immunoassay and evaluated the clinical accuracy of the method.MethodsA total of 90 patients with chest pain admitted to Wuxi Second People’s Hospital of Nanjing Medical University had peripheral blood and venous blood samples collected for detection of hs-cTnI by rapid quantum dot fluorescence immunoassay. The differences between the two methods were evaluated, as well as the analytical performance and clinical diagnostic efficacy of hs-cTnI detection by quantum dot fluorescence immunoassay. The final diagnosis was determined by two independent cardiologists.ResultsThis study verified the precision, linear range and sensitivity of the novel detection method. There was good correlation between the results of hs-cTnI quantum dot fluorescence immunoassay for peripheral blood and the results for venous blood (regression equation Y=1.026x+0.521, R2=0.9337); 94.4% (85/90) of the data were within the conformance limit. In addition, in the analysis of 52 patients with confirmed AMI, the clinical specificity of the quantum dot fluorescence immunoassay in peripheral blood was the same as that in venous blood samples (89.5%:89.5%). Finally, the area under the receiver operating characteristic (ROC) curve of the peripheral blood quantum dot fluorescence immunoassay was 0.9352, the 95% confidence interval (CI) was 0.8829 to 0.9876, the cut-off value was 1.598, and the sensitivity was 82.69%, which was not significantly different from the venous blood method (P value =0.089).ConclusionsRapid detection of hs-cTnI by quantum dot fluorescence immunoassay in peripheral blood is feasible. It has a high correlation and consistency with the venous blood method, as well as a high clinical diagnostic value for AMI and is more convenient and easier to detect.

T cells continuously sample CNS-derived antigens in the periphery, yet it is unknown how they sample and respond to CNS antigens derived from distinct brain areas. We expressed ovalbumin (OVA) neoepitopes in regionally distinct CNS areas (Cnp-OVA and Nes-OVA mice) to test peripheral antigen sampling by OVA-specific T cells under homeostatic and neuroinflammatory conditions. We show that antigen sampling in the periphery is independent of regional origin of CNS antigens in both male and female mice. However, experimental autoimmune encephalomyelitis (EAE) is differentially influenced in Cnp-OVA and Nes-OVA female mice. Although there is the same frequency of CD45high CD11b+ CD11c+ CX3CL1+ myeloid cell-T-cell clusters in neoepitope-expressing areas, EAE is inhibited in Nes-OVA female mice and accelerated in CNP-OVA female mice. Accumulation of OVA-specific T cells and their immunomodulatory effects on EAE are CX3C chemokine receptor 1 (CX3CR1) dependent. These data show that despite similar levels of peripheral antigen sampling, CNS antigen-specific T cells differentially influence neuroinflammatory disease depending on the location of cognate antigens and the presence of CX3CL1/CX3CR1 signaling.SIGNIFICANCE STATEMENT Our data show that peripheral T cells similarly recognize neoepitopes independent of their origin within the CNS under homeostatic conditions. Contrastingly, during ongoing autoimmune neuroinflammation, neoepitope-specific T cells differentially influence clinical score and pathology based on the CNS regional location of the neoepitopes in a CX3CR1-dependent manner. Altogether, we propose a novel mechanism for how T cells respond to regionally distinct CNS derived antigens and contribute to CNS autoimmune pathology.

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MicroRNA-21 (miR-21) contributes to the pathogenesis of fibrogenic diseases in multiple organs, including the kidneys, potentially by silencing metabolic pathways that are critical for cellular ATP generation, ROS production, and inflammatory signaling. Here, we developed highly specific oligonucleotides that distribute to the kidney and inhibit miR-21 function when administered subcutaneously and evaluated the therapeutic potential of these anti-miR-21 oligonucleotides in chronic kidney disease. In a murine model of Alport nephropathy, miR-21 silencing did not produce any adverse effects and resulted in substantially milder kidney disease, with minimal albuminuria and dysfunction, compared with vehicle-treated mice. miR-21 silencing dramatically improved survival of Alport mice and reduced histological end points, including glomerulosclerosis, interstitial fibrosis, tubular injury, and inflammation. Anti-miR-21 enhanced PPARα/retinoid X receptor (PPARα/RXR) activity and downstream signaling pathways in glomerular, tubular, and interstitial cells. Moreover, miR-21 silencing enhanced mitochondrial function, which reduced mitochondrial ROS production and thus preserved tubular functions. Inhibition of miR-21 was protective against TGF-β-induced fibrogenesis and inflammation in glomerular and interstitial cells, likely as the result of enhanced PPARα/RXR activity and improved mitochondrial function. Together, these results demonstrate that inhibition of miR-21 represents a potential therapeutic strategy for chronic kidney diseases including Alport nephropathy.