概要: Because epithelial surface pH affects airway inflammation and host defense, we study the benefit of an inhaled buffer in patients with asthma. Inhaled Optate is designed safely to raise airway surface pH and is well-tolerated in humans. Epithelial infection with SARS-CoV2 is attenuated by raising intracellular pH. Here, we show that Optate raises intracellular pH in primary normal human airway epithelial (NHAE) cells without affecting cell viability. Moreover, doses of Optate used in our human trials inhibit SARS-CoV-2 replication in NHAE cells. Specifically, Optate almost completely ablates viral plaque formation and nucleocapsid protein expression, decreasing expression of critical processing proteins. Optate could be a candidate to evaluate for treating COVID-19 patients.
项目整体设计: Six samples of extracted and purified RNA from one line of primary human airway epithelial cells were processed for RNAseq analysis. This included three samples that were exposed to Optate for 48 hours and three control samples.
1 X 10^5 cells (second passage) were plated and allowed to adhere to the culture plate overnight. These cells underwent 48 hours of treatment (or control) and then underwent RNA extraction; both treatment and extraction are described below.
处理方案:
-
提取方案:
Cells were lysed and RNA was extracted using the Qiagen RNeasy Plus Mini Kit per manufacturers protocol (Qiagen, USA, Cat No. 74134).
建库方案:
-
测序信息
分子类型:
poly(A)+ RNA
库的片段类型:
SINGLE
库的链类型:
Forward
测序平台:
ILLUMINA
测序仪型号:
Illumina NextSeq 500
链特异性:
Specific
样本
基本信息:
样本描述:
生物条件:
实验变量:
方案:
测序信息:
质量评估:
数据来源
GEN样本编号
GEN数据集编号
系列编号
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样本编号
样本名称
生物样本编号
样本访问号
实验访问号
释放时间
提交时间
最后更新时间
物种
种族
族裔
年龄
年龄单位
性别
来源名称
组织
细胞类型
细胞亚型
细胞系
疾病
疾病状态
发育阶段
突变/变异
表型
Condition Detail
生长方案
处理方案
提取方案
建库方案
分子类型
库的片段类型
链特异性
库的链类型
加标（Spike-In）
测序方法
测序平台
测序仪型号
细胞数
测序片段数
碱基数
平均测序片段长度_1
平均测序片段长度_2
唯一比对率
多重比对率
覆盖度
文章
A Treatment to Eliminate SARS-CoV-2 Replication in Human Airway Epithelial Cells Is Safe for Inhalation as an Aerosol in Healthy Human Subjects.
