Targeting Intra-Pulmonary P53-Dependent Long Non-Coding RNA Expression as a Therapeutic Intervention for Systemic Lupus Erythematosus-Associated Diffuse Alveolar Hemorrhage

Abstract

Diffuse alveolar hemorrhage (DAH) in systemic lupus erythematosus (SLE) is associated with significant mortality, requiring a thorough understanding of its complex mechanisms to develop novel therapeutics for disease control. Activated p53-dependent apoptosis with dysregulated long non-coding RNA (lncRNA) expression is involved in the SLE pathogenesis and correlated with clinical activity. We examined the expression of apoptosis-related p53-dependent lncRNA, including H19, HOTAIR and lincRNA-p21 in SLE-associated DAH patients. Increased lincRNA-p21 levels were detected in circulating mononuclear cells, mainly in CD4+ and CD14+ cells. Higher expression of p53, lincRNA-p21 and cell apoptosis was identified in lung tissues. Lentivirus-based short hairpin RNA (shRNA)-transduced stable transfectants were created for examining the targeting efficacy in lncRNA. Under pristane stimulation, alveolar epithelial cells had increased p53, lincRNA-p21 and downstream Bax levels with elevated apoptotic ratios. After pristane injection, C57/BL6 mice developed DAH with increased pulmonary expression of p53, lincRNA-p21 and cell apoptosis. Intra-pulmonary delivery of shRNA targeting lincRNA-p21 reduced hemorrhage frequencies and improved anemia status through decreasing Bax expression and cell apoptosis. Our findings demonstrate increased p53-dependent lncRNA expression with accelerated cell apoptosis in the lungs of SLE-associated DAH patients, and show the therapeutic potential of targeting intra-pulmonary lncRNA expression in a pristane-induced model of DAH.

Identification of Diagnostic Biomarkers in Systemic Lupus Erythematosus Based on Bioinformatics Analysis and Machine Learning

“…PRIC285 is a transcriptional coactivator involved in PPAR-γ signaling ( Fairfax et al, 2012 ), and PPAR-γ can inhibit the activation of macrophages and regulate their differentiation, improving SLE symptoms ( Kiss et al, 2013 ). XAF1 can increase p53 transcriptional activity ( Pinto et al, 2020 ), and p53-dependent apoptosis has been implicated in the pathogenesis and disease activity of SLE ( Chen et al, 2021 ).…”

Long non‐coding RNAs in systemic lupus erythematosus: New insights into disease pathogenesis and diagnosis

“…Moreover, intra‐pulmonary delivery of shRNA targeting lincRNA‐p21 could decrease haemorrhage and improve anaemia through reducing cell apoptosis in the lungs of a mouse model. These studies indicate the therapeutic potential of targeting lincRNA‐p21 in clinical patients 76 …”

Down-regulated miR-146a expression with increased neutrophil extracellular traps and apoptosis formation in autoimmune-mediated diffuse alveolar hemorrhage

“…We further created stable LV-sh-miR-146 #1-transfected transfectants by using puromycin selection process in sorted CRISPR-CasRX-02-transfected HL-60 cells (Additional file 1: Fig. S1c, for miR-146a, sh-luciferase versus sh-miR-146a, 100.0 ± 3.8 versus 19.2 ± 1.4%, p = 0.003) [5].…”

“…Total RNAs from formalin-fixed, paraffin-embedded human tissues were purified by RNeasy FFPE Kit (Qiagen, Hilden, Germany with hydrophilic surface, to effectively deliver pristane in vitro [22]. A bolus of 2 mM pristane (Sigma-Aldrich) was pipetted into 4 mM solution of βCD (Sigma-Aldrich) and stirred at RT for 4 d [5]. The crystalline complexes which precipitated out of solution were washed twice in PBS and stored at 4 °C [22].…”

“…Overactivated neutrophils with neutrophil extracellular traps (NETs) formation and impaired removal, contributes to disease development and progression by providing autoantigens and increasing proinflammatory responses. Enhanced apoptosis and NETs formation have been demonstrated in the SLE-mediated DAH lungs [5,6].…”