Poly(L-glutamic acid)-co-poly(ethylene glycol) block copolymers for protein conjugation

Poly(L-glutamic acid)-co-poly(ethylene glycol) block copolymers (PLE-PEG) are here investigated as polymers for conjugation to therapeutic proteins such as granulocyte colony stimulating factor (G-CSF) and human growth hormone (hGH). PLE-PEG block copolymers are able to stabilize and protect proteins from degradation and to prolong their residence time in the blood stream, features that are made possible thanks to PEG's intrinsic properties and the simultaneous presence of the biodegradable anionic PLE moiety. When PLE-PEG copolymers are selectively tethered to the N-terminus of G-CSF and hGH, they yield homogeneous monoconjugates that preserve the protein's secondary structure. During the current study the pharmacokinetics of PLE10-PEG20k-G-CSF and PLE20-PEG20k-G-CSF derivatives and their ability to induce granulopoiesis were, respectively, assessed in Sprague-Dawley rats and in C57BL6 mice. Our results show that the bioavailability and bioactivity of the derivatives are comparable to or better than those of PEG20k-Nter-G-CSF (commercially known as Pegfilgrastim). The therapeutic effects of PLE10-PEG20k-hGH and PLE20-PEG20k-hGH derivatives tested in hypophysectomized rats demonstrate that the presence of a negatively charged PLE block enhances the biological properties of the conjugates additionally with respect to PEG20k-Nter-hGH

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

Correlation between movement complexity during static standing and balance function in institutionalized older adults

Purpose: Sample entropy (SampEn) is an analysis to evaluate movement complexity of the center of pressure (COP). A lower value of SampEn indicates lower complexity of COP variability, that is, rigidity, and lower degrees of freedom. Previous studies reported the association of increased SampEn with improved standing balance ability in young subjects. However, no studies have examined these relationships among older adults. Thus, we aimed to investigate the relationship between SampEn and standing balance ability in older adults. Subjects and methods: The subjects were 33 institutionalized older adults (aged 82.2±6.5 years). COP during static standing was measured. The standard deviation (SD) values of COP and SampEn in the sagittal and frontal planes were calculated using time series data. One-leg standing test (OLST), functional reach (FR) test, and lateral reach (LR) test were also measured to evaluate standing balance ability. Results: OLST, FR, and LR were 6.5±8.3 s, 19.8±5.9 cm, and 18.2±6.4 cm, respectively. Pearson correlation analysis revealed that SampEn in the sagittal plane significantly correlated with OLST (r=-0.35) and FR (r=-0.36). However, SampEn in the frontal plane and SD of COP in both sagittal and frontal planes had no relationship with any of the clinical balance tests. Conclusion: Lower SampEn implies rigidity for postural control. In the present study, it was found that lower SampEn in the sagittal plane was related to a higher balance function, which suggests that older adults utilized body rigidity to maintain postural stability as a compensative strategy

Association of activities of daily living with the load during step ascent motion in nursing home-residing elderly individuals

Objective: This study aimed to examine the association of independence in activities of daily living with the loads during step ascent motion and other motor functions in 32 nursing home–residing elderly individuals. Design: Independence in activities of daily living was assessed by using the Functional Independence Measure. The loads at the upper (i.e., pulling up) and lower (i.e., pushing up) levels during the step ascent task was measured on a step ascent platform. Hip extensor, knee extensor, plantar flexor muscle, and quadriceps setting strengths; lower extremity agility using the stepping test; and hip and knee joint pain severities were measured. One-legged stance and functional reach distance for balance and maximal walking speed, timed up-and-go time, five-chair-stand time, and step ascent time were also measured to assess mobility. Results: Stepwise regression analysis revealed that the load at pushing up during step ascent motion and timed up-and-go time were significant and independent determinants of Functional Independence Measure score. Functional Independence Measure score decreased with decreased load at pushing up and increased timed up-and-go time. Conclusions: The study results suggest that depending on task specificity, both one step up task's push-up peak load during step ascent motion and timed up-and-go can partially explain activities of daily living's Functional Independence Measure score in nursing home–residing elderly individuals. Lower extremity muscle strength, agility, pain, or balance measures did not add to the prediction