Role of Dok-1 and Dok-2 in Myeloid Homeostasis and Suppression of Leukemia

Dok-1 and Dok-2 are closely related rasGAP-associated docking proteins expressed preferentially in hematopoietic cells. Although they are phosphorylated upon activation of many protein tyrosine kinases (PTKs), including those coupled with cytokine receptors and oncogenic PTKs like Bcr-Abl, their physiological roles are largely unidentified. Here, we generated mice lacking Dok-1 and/or Dok-2, which included the double-deficient mice succumbed to myeloproliferative disease resembling human chronic myelogenous leukemia (CML) and chronic myelomonocytic leukemia. The double-deficient mice displayed medullary and extramedullary hyperplasia of granulocyte/macrophage progenitors with leukemic potential, and their myeloid cells showed hyperproliferation and hypo-apoptosis upon treatment and deprivation of cytokines, respectively. Consistently, the mutant myeloid cells showed enhanced Erk and Akt activation upon cytokine stimulation. Moreover, loss of Dok-1 and/or Dok-2 induced blastic transformation of chronic phase CML-like disease in mice carrying the bcr-abl gene, a cause of CML. These findings demonstrate that Dok-1 and Dok-2 are key negative regulators of cytokine responses and are essential for myeloid homeostasis and suppression of leukemia

A de novo dominant-negative variant is associated with OTULIN-related autoinflammatory syndrome

稀少遺伝性自己炎症性疾患: OTULIN関連自己炎症症候群の新たな病態を解明～既報の疾患に新たな視点を追加し、未診断患者の診断や炎症・細胞死研究の進展に期待～. 京都大学プレスリリース. 2024-04-25.OTULIN-related autoinflammatory syndrome (ORAS), a severe autoinflammatory disease, is caused by biallelic pathogenic variants of OTULIN, a linear ubiquitin-specific deubiquitinating enzyme. Loss of OTULIN attenuates linear ubiquitination by inhibiting the linear ubiquitin chain assembly complex (LUBAC). Here, we report a patient who harbors two rare heterozygous variants of OTULIN (p.P152L and p.R306Q). We demonstrated accumulation of linear ubiquitin chains upon TNF stimulation and augmented TNF-induced cell death in mesenchymal stem cells differentiated from patient-derived iPS cells, which confirms that the patient has ORAS. However, although the de novo p.R306Q variant exhibits attenuated deubiquitination activity without reducing the amount of OTULIN, the deubiquitination activity of the p.P152L variant inherited from the mother was equivalent to that of the wild-type. Patient-derived MSCs in which the p.P152L variant was replaced with wild-type also exhibited augmented TNF-induced cell death and accumulation of linear chains. The finding that ORAS can be caused by a dominant-negative p.R306Q variant of OTULIN furthers our understanding of disease pathogenesis

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

Elderly patients aged over 75 years with glioblastoma: Preoperative status and surgical strategies

Background: Standards of care for the elderly with glioblastoma are still unknown, because most studies exclude elderly patients. The effects of preoperative status and surgical strategies on the prognosis for elderly glioblastoma patients were studied. Methods: We defined “elderly” as 75 years or older and retrospectively reviewed 137 patients with glioblastoma, including 31 (22.6%) elderly patients. Relationships between age and clinicopathological variables were investigated. Results: The median overall survival (OS) was 15.8 months in the nonelderly group and 10.8 months in the elderly group (p = 0.02). The median progression-free survival (PFS) was 9.1 months in the nonelderly group and 6.6 months in the elderly group (p = 0.02). Median OS and median PFS had no relationship between low (<70) and high (≥70) Karnofsky performance status (KPS) in the nonelderly group. However, the median OS was significantly longer in the elderly group with high KPS (8.4 months in low KPS, 12.4 months in high KPS; p = 0.003). The median PFS was also significantly longer in the elderly group with high KPS (4.3 months in low KPS, 9.1 months in high KPS; p = 0.04). OS and PFS were significantly longer in the nonelderly group with resection than with biopsy (OS, p = 0.016; PFS, p = 0.039). However, neither OS nor PFS showed any difference with surgical method in the elderly group (OS, p = 0.241; PFS, p = 0.131). Conclusions: Less invasive treatment can be considered as a treatment option in addition to radical resection in glioblastoma patients aged 75 years and older with KPS on admission of less than 70, depending on the general condition

Self-diagnosis of seasonal influenza in a rural primary care setting in Japan: A cross sectional observational study

<div><p>Objective</p><p>To elucidate the accuracy and optimal cut-off point of self-diagnosis and clinical symptoms of seasonal influenza compared with rapid influenza diagnostic tests as the reference standard, we conducted a cross sectional observational study at a rural clinic in Japan.</p><p>Methods</p><p>Data during three influenza seasons (December 2013 to April 2016) were retrospectively collected from the medical records and pre-examination sheets of 111 patients aged >11 years (mean age 48.1 years, men 53.2%) who were suspected of influenza infection and underwent rapid influenza diagnostic testing. Patients’ characteristics (age, sex, and past medical history of influenza infection), clinical signs (axillary temperature, pulse rate, cough, joint and muscle pain, and history of fever [acute or sudden, gradual, and absence of fever]), duration from the onset of symptoms, severity of feeling sick compared with a common cold (severe, similar, and mild), self-reported likelihood of influenza (%), and results of rapid influenza diagnostic tests.</p><p>Results</p><p>At the optimal cut-off point (30%) for estimation of self-diagnosis of seasonal influenza, the positive likelihood ratio (LR+) was 1.46 (95% confidence interval 1.07 to 2.00) and negative likelihood ratio (LR–) was 0.57 (0.35 to 0.93). At a 10% cut-off point, LR–was 0.33 (0.12 to 0.96). At an 80% cut-off point, LR+ was 2.75 (0.75 to 10.07). As for clinical signs, the combination of acute or sudden onset fever and cough had LR+ of 3.27 (1.68 to 6.35). Absence of cough showed LR–of 0.15 (0.04 to 0.61).</p><p>Conclusions</p><p>Self-diagnosis of influenza using the optimal cut-off point (30%) was not found useful for ruling in or ruling out an influenza diagnosis. However, it could be useful when patients self-report extremely high (80%) or low (10%) probability of having influenza. Clinically useful signs were the combination of history of fever and cough, and absence of cough.</p></div