CRISPRi screens identify the lncRNA, LOUP, as a multifunctional locus regulating macrophage differentiation and inflammatory signaling.

Long noncoding RNAs (lncRNAs) account for the largest portion of RNA from the transcriptome, yet most of their functions remain unknown. Here, we performed two independent high-throughput CRISPRi screens to understand the role of lncRNAs in monocyte function and differentiation. The first was a reporter-based screen to identify lncRNAs that regulate TLR4-NFkB signaling in human monocytes and the second screen identified lncRNAs involved in monocyte to macrophage differentiation. We successfully identified numerous noncoding and protein-coding genes that can positively or negatively regulate inflammation and differentiation. To understand the functional roles of lncRNAs in both processes, we chose to further study the lncRNA LOUP [lncRNA originating from upstream regulatory element of SPI1 (also known as PU.1)], as it emerged as a top hit in both screens. Not only does LOUP regulate its neighboring gene, the myeloid fate-determining factor SPI1, thereby affecting monocyte to macrophage differentiation, but knockdown of LOUP leads to a broad upregulation of NFkB-targeted genes at baseline and upon TLR4-NFkB activation. LOUP also harbors three small open reading frames capable of being translated and are responsible for LOUP's ability to negatively regulate TLR4/NFkB signaling. This work emphasizes the value of high-throughput screening to rapidly identify functional lncRNAs in the innate immune system.

Combination Therapy with KRAS and P38α siRNA Suppresses Colorectal Cancer Growth and Development in SW480 Cell Line.

PURPOSE: Colorectal cancer (CRC) is one of the most prevalence malignancies in a different society with a high rate of death. The KRAS and p38α axes have critical roles in the development, migration, and growth of numerous tumors, such as colorectal malignancy. KRAS mutation acts as an oncogene in various cancers and is correlated with the poor prognosis in colorectal tumors. Also, p38α plays different roles and exhibits tissue-dependent activity. In some tissues act as an oncogene while in others act as a tumor suppressor. In this research, we try to understand the effect of the P38α and KRAS genes suppression by specific siRNAs on the SW480 cell line progression. METHODS: We evaluate the impact of the P38α and KRAS gene knockdown by special siRNA on the growth and development of the SW480 cell line. SW480 cell line was treated with KRAS and P38α siRNAs, and the cell viability, gene expression, migration ability, and rate of apoptosis were evaluated with MTT assay, real-time PCR, scratch test, and flow cytometry. RESULTS: After treatment of the cancer cell with KRAs and P38α siRNAs, cell viability reduced to 29.16%. Also, the expression levels of the KRAS and P38α genes reduced to 26.34% and 16.06%, respectively. Apoptosis rate after combination therapy with KRAS and P38α siRNAs increased to 72.1. Also, we found that these siRNAs suppress cell migration in SW480 cell lines. CONCLUSION: The current study showed that combination therapy with p38α and KRAS siRNA may be considered a novel therapy for colorectal tumor in future.

Relationship between chest CT scan findings with SOFA score, CRP, comorbidity, and mortality in ICU patients with COVID-19.

OBJECTIVE: This study aimed to investigate the relationship between chest computed tomography (CT) scan findings with sequential organ failure assessment (SOFA) score, C-reactive protein (CRP), comorbidity, and mortality in intensive care unit (ICU) patients with coronavirus disease 19 (COVID-19). METHOD: Adult patients (≥18 years old) with COVID-19 who were consecutively admitted to the Imam-Reza Hospital, Tabriz, East-Azerbaijan Province, North-West of Iran between March 2020 and August 2020 were screened and total of 168 patients were included. Demographic, clinical, and mortality data were gathered. Severity of disease was evaluated using the SOFA score system. CRP levels were measured and chest CT scans were performed. RESULTS: Most of patients had multifocal and bilateral ground glass opacity (GGO) pattern in chest CT scan. There were significant correlations between SOFA score on admission with multifocal and bilateral GGO (P = .010 and P = .011, respectively). Significant relationships were observed between unilateral and bilateral GGO patterns with CRP (P = .049 and P = .046, respectively). There was significant relationship between GGO patterns with comorbidities including overweight/obesity, heart failure, cardiovascular diseases, and malignancy (P < .05). No significant relationships were observed between chest CT scan results with mortality (P > .05). CONCLUSION: Multifocal bilateral GGO was the most common pattern. Although chest CT scan characteristics were significantly related with SOFA score, CRP, and comorbidity in ICU patients with COVID-19, a relationship with mortality was not significant.

Lack of association between six-minute walk distance and pulmonary artery pressure in patients with systemic sclerosis and cardiopulmonary symptoms.

The six-minute walk test (6MWT) is a non-invasive test used to assess cardiopulmonary performance. The aim of this study was to evaluate the performance of 6MWT in predicting pulmonary artery hypertension (PAH) and interstitial lung disease in patients with systemic sclerosis (SSc) and cardiopulmonary symptoms. Sixty-three patients with SSc who had dyspnea, cough, chest pain and syncope underwent 6MWT, high-resolution computed tomography (HRCT), spirometry, body plethysmography and single breath carbon monoxide diffusion measurement. There were no significant differences in mean 6MWD between patients with diffuse SSc compared with limited disease, patients with no parenchymal involvement compared with patients with parenchymal involvement <20% and≥20% in HRCT, and patients with PAP ≥25 mm Hg compared with patents with PAP <25 mm Hg. No significant relationship was found between 6MWD and age, mean PAP, forced expiratory volume, forced vital capacity and diffusing capacity of the lungs for carbon monoxide. The present study showed that in patients with SSc and cardiopulmonary symptoms, 6MVT does not help to predict PAH and parenchymal lung involvement.