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Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion.

单细胞糖酵解活性调节膜张力和 HIV-1 融合。

  • 影响因子:6.02
  • DOI:10.1371/journal.ppat.1008359
  • 作者列表:"Coomer CA","Carlon-Andres I","Iliopoulou M","Dustin ML","Compeer EB","Compton AA","Padilla-Parra S
  • 发表时间:2020-02-21
Abstract

:There has been resurgence in determining the role of host metabolism in viral infection yet deciphering how the metabolic state of single cells affects viral entry and fusion remains unknown. Here, we have developed a novel assay multiplexing genetically-encoded biosensors with single virus tracking (SVT) to evaluate the influence of global metabolic processes on the success rate of virus entry in single cells. We found that cells with a lower ATP:ADP ratio prior to virus addition were less permissive to virus fusion and infection. These results indicated a relationship between host metabolic state and the likelihood for virus-cell fusion to occur. SVT revealed that HIV-1 virions were arrested at hemifusion in glycolytically-inactive cells. Interestingly, cells acutely treated with glycolysis inhibitor 2-deoxyglucose (2-DG) become resistant to virus infection and also display less surface membrane cholesterol. Addition of cholesterol in these in glycolytically-inactive cells rescued the virus entry block at hemifusion and enabled completion of HIV-1 fusion. Further investigation with FRET-based membrane tension and membrane order reporters revealed a link between host cell glycolytic activity and host membrane order and tension. Indeed, cells treated with 2-DG possessed lower plasma membrane lipid order and higher tension values, respectively. Our novel imaging approach that combines lifetime imaging (FLIM) and SVT revealed not only changes in plasma membrane tension at the point of viral fusion, but also that HIV is less likely to enter cells at areas of higher membrane tension. We therefore have identified a connection between host cell glycolytic activity and membrane tension that influences HIV-1 fusion in real-time at the single-virus fusion level in live cells.

摘要

: 在确定宿主代谢在病毒感染中的作用方面已经复苏,但破译单细胞的代谢状态如何影响病毒进入和融合仍然未知。在此,我们开发了一种新型的检测复用基因编码的生物传感器与单一病毒跟踪 (SVT),以评估全球代谢过程对单细胞中病毒进入成功率的影响。我们发现在加入病毒之前 ATP: ADP 比值较低的细胞对病毒融合和感染的许可程度较低。这些结果表明宿主代谢状态与病毒-细胞融合发生的可能性之间存在关系。SVT 显示,HIV-1 病毒体在半融合时在糖代谢不活跃的细胞中被阻滞。有趣的是,糖酵解抑制剂 2-脱氧葡萄糖 (2-DG) 急性处理的细胞对病毒感染具有抵抗力,并且还显示出较少的表面膜胆固醇。在糖酵解不活跃的细胞中加入胆固醇可挽救半融合时的病毒进入阻滞,并使 HIV-1 融合得以完成。进一步研究基于 FRET 的膜张力和膜顺序报告者揭示了宿主细胞糖酵解活性与宿主膜顺序和张力之间的联系。事实上,用 2-DG 处理的细胞分别具有较低的质膜脂质顺序和较高的张力值。我们结合终身成像 (FLIM) 和 SVT 的新型成像方法不仅揭示了病毒融合点质膜张力的变化, 而且,艾滋病毒不太可能进入膜张力较高的区域的细胞。因此,我们已经确定了宿主细胞糖酵解活性和膜张力之间的联系,在活细胞的单病毒融合水平上实时影响 HIV-1 融合。

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DOI:10.1111/ajo.13112
作者列表:["Alhayo S","Leonardi M","Lu C","Gosal P","Reid S","Barto W","Condous G"]

METHODS:BACKGROUND:Ultrasound has been demonstrated to accurately diagnose rectal deep endometriosis (DE) and pouch of Douglas (POD) obliteration. The role of ultrasound in the assessment of patients who have undergone surgery for rectal DE and POD obliteration has not been evaluated. AIM:To describe the transvaginal ultrasound (TVS) findings of patients who have undergone rectal surgery for DE. MATERIALS AND METHODS:An observational cross-sectional study at a tertiary care centre in Sydney, Australia between January and April 2017. Patients previously treated for rectal DE (low anterior resection vs rectal shaving/disc excision) were recruited and asked to complete a questionnaire on their current symptoms. On TVS, POD state and rectal DE were assessed. Correlating recurrence of POD obliteration and/or rectal DE to surgery type and symptoms was done. RESULTS:Fifty-six patients were contacted; 22/56 (39.3%) attended for the study visit. Average interval of surgery to study visit was 52.8 ± 24.6 months. Surgery type breakdown was as follows: low anterior resection (56%) and rectal shaving/disc excision (44%). The prevalence of POD obliteration was 16/22 (72.7%) intraoperatively and 8/22 (36.4%) at study visit, as per the sliding sign. Nine patients (39.1%) had evidence on TVS of recurrent rectal DE. Recurrence of POD obliteration and rectal DE was not associated with surgery type or symptomatology. CONCLUSION:Despite surgery for rectal DE, many patients have a negative sliding sign on TVS, representing POD obliteration, and rectal DE. Our numbers are too small to correlate with the surgery type or their current symptoms.

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发表时间:2020-01-10
DOI:10.1007/s00261-019-02399-0
作者列表:["Burnett TL","Feldman MK","Huang JQ"]

METHODS::Minimally invasive surgery for complex endometriosis requires preoperative planning that intimately connects the gynecologic surgeon to the radiologist. Understanding the surgeon's perspective to endometriosis treatment facilitates a productive relationship that ultimately benefits the patient. We examine minimally invasive surgery for endometriosis and the key radiologic information which enable the surgeon to successfully negotiate patient counseling, preoperative planning, and an interdisciplinary approach to surgery.

影响因子:1.69
发表时间:2020-01-09
DOI:10.1016/j.jmig.2020.01.002
作者列表:["Byun J","Peterson CM","Backonja U","Taylor RN","Stanford JB","Allen-Brady KL","Smith KR","Buck Louis GM","Schliep KC"]

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