Exploring the role of microRNAs in airway smooth muscle biology and asthma therapy

The pathophysiology of asthma is characterized by airway inflammation, remodeling and hyper-responsiveness. Phenotypic changes in airway smooth muscle cells (ASM) play a pivotal role in the pathogenesis of asthma. ASM cells promote inflammation and are key drivers of airway remodeling. While airway hyper responsiveness and inflammation can be managed by bronchodilators and anti-inflammatory drugs, ASM remodeling is poorly managed by existing therapies. Therefore, targeting ASM remodeling remains a challenge, and a deeper understanding of the molecular mechanism that regulates ASM phenotypes in asthma pathogenesis will facilitate the search for next-generation asthma therapy. MicroRNAs are small yet versatile gene tuners that regulate a variety of cellular processes, including cell proliferation and inflammation - two phenotypes that are often altered in asthmatic ASM. We thus hypothesized that microRNAs regulate ASM phenotypes in asthma and represent new targets for future therapy. In this thesis, we used a genomic approach that combined next-generation sequencing with functional cellular assays to characterize the role of microRNAs in regulating airway smooth muscle function and drug response to conventional therapies. In Chapter 2, we identified miR-10a as the most abundant microRNA expressed in the primary human airway smooth muscle (HASM) cells. Using an unbiased target identification approach, we identified several novel potential targets of miR-10a, including the catalytic subunit alpha of PI3 kinase (PIK3CA)--the central component of the PI3K pathway. We demonstrated that miR-10a directly suppresses PIK3CA expression by targeting its 3' Untranslated region (3'-UTR). Inhibition of PIK3CA by miR-10a reduced AKT phosphorylation and blunted the expression of cyclins and cyclin-dependent kinases that are required for HASM proliferation. In Chapter 3, we examined the effect of conventional asthma therapies on miRNA expression. While we did not find significant changes in miRNA levels, it remains to be determined whether microRNAs play a role in ASM tissue response to asthma therapy. Our study is the first to examine the role of microRNAs in ASM proliferation. Results from our study identified a novel microRNA-mediated regulatory mechanism of PI3K signaling and ASM proliferation. They suggest further that miR-10a is a potential therapeutic target to treat airway remodeling in asthma

A Duty to Forget, a Right to be Assured? Exposing Vulnerabilities in Machine Unlearning Services

The right to be forgotten requires the removal or "unlearning" of a user's data from machine learning models. However, in the context of Machine Learning as a Service (MLaaS), retraining a model from scratch to fulfill the unlearning request is impractical due to the lack of training data on the service provider's side (the server). Furthermore, approximate unlearning further embraces a complex trade-off between utility (model performance) and privacy (unlearning performance). In this paper, we try to explore the potential threats posed by unlearning services in MLaaS, specifically over-unlearning, where more information is unlearned than expected. We propose two strategies that leverage over-unlearning to measure the impact on the trade-off balancing, under black-box access settings, in which the existing machine unlearning attacks are not applicable. The effectiveness of these strategies is evaluated through extensive experiments on benchmark datasets, across various model architectures and representative unlearning approaches. Results indicate significant potential for both strategies to undermine model efficacy in unlearning scenarios. This study uncovers an underexplored gap between unlearning and contemporary MLaaS, highlighting the need for careful considerations in balancing data unlearning, model utility, and security.Comment: To Appear in the Network and Distributed System Security Symposium (NDSS) 2024, San Diego, CA, US

How government green investment affects the carbon emission reduction process: empirical evidence from prefecture-level cities in China

Government green investment (GGI) is one of the effective tools for reducing carbon emissions (CEs). This is of great significance for the realization of “carbon peaking and carbon neutrality.” This study innovatively considers the multidimensional CE reduction (CER) process indexes to explore the impact mechanism of GGI on China’s CER process. At the same time, CER is particularly critical in resource-dependent regions. This study incorporates this perspective to explore the CER effect of GGI in these regions. This paper developed a multidimensional evaluation system for China’s CER process, using panel data of 269 prefecture-level cities from 2008 to 2019 to explore the impact of GGI on China’s CER process. The results indicated that 1) GGI promotes CER in China as a whole and effectively inhibits CEs, per capita CEs, and CE intensity; 2) GGI promotes CER to some extent by enhancing the energy efficiency and total factor productivity; 3) it plays a larger role in CER in regions with a high energy endowment; and 4) the impact of GGI on CER is heterogeneous in geographical regions, city sizes, and economic development levels. This study makes policy recommendations for reducing CEs, including intensifying GGI and playing its investment-pulling role, thereby increasing the investment related to improving energy efficiency and total factor productivity and promoting government intervention in areas with high energy endowments

Low-velocity-favored transition radiation

When a charged particle penetrates through an optical interface, photon emissions emerge - a phenomenon known as transition radiation. Being paramount to fundamental physics, transition radiation has enabled many applications from high-energy particle identification to novel light sources. A rule of thumb in transition radiation is that the radiation intensity generally decreases with the particle velocity v; as a result, low-energy particles are not favored in practice. Here we find that there exist situations where transition radiation from particles with extremely low velocities (e.g. v/c<0.001) exhibits comparable intensity as that from high-energy particles (e.g. v/c=0.999), where c is light speed in free space. The comparable radiation intensity implies an extremely high photon extraction efficiency from low-energy particles, up to eight orders of magnitude larger than that from high-energy particles. This exotic phenomenon of low-velocity-favored transition radiation originates from the excitation of Ferrell-Berreman modes in epsilon-near-zero materials. Our findings may provide a promising route towards the design of integrated light sources based on low-energy electrons and specialized detectors for beyond-standard-model particles.Comment: 13 pages, 4 figure

RNA-Seq Transcriptome Profiling Identifies CRISPLD2 as a Glucocorticoid Responsive Gene that Modulates Cytokine Function in Airway Smooth Muscle Cells

Asthma is a chronic inflammatory respiratory disease that affects over 300 million people worldwide. Glucocorticoids are a mainstay therapy for asthma because they exert anti-inflammatory effects in multiple lung tissues, including the airway smooth muscle (ASM). However, the mechanism by which glucocorticoids suppress inflammation in ASM remains poorly understood. Using RNA-Seq, a high-throughput sequencing method, we characterized transcriptomic changes in four primary human ASM cell lines that were treated with dexamethasone—a potent synthetic glucocorticoid (1 µM for 18 hours). Based on a Benjamini-Hochberg corrected p-value <0.05, we identified 316 differentially expressed genes, including both well known (DUSP1, KLF15, PER1, TSC22D3) and less investigated (C7, CCDC69, CRISPLD2) glucocorticoid-responsive genes. CRISPLD2, which encodes a secreted protein previously implicated in lung development and endotoxin regulation, was found to have SNPs that were moderately associated with inhaled corticosteroid resistance and bronchodilator response among asthma patients in two previously conducted genome-wide association studies. Quantitative RT-PCR and Western blotting showed that dexamethasone treatment significantly increased CRISPLD2 mRNA and protein expression in ASM cells. CRISPLD2 expression was also induced by the inflammatory cytokine IL1β, and small interfering RNA-mediated knockdown of CRISPLD2 further increased IL1β-induced expression of IL6 and IL8. Our findings offer a comprehensive view of the effect of a glucocorticoid on the ASM transcriptome and identify CRISPLD2 as an asthma pharmacogenetics candidate gene that regulates anti-inflammatory effects of glucocorticoids in the ASM

Elevated limb-bud and heart development (LBH) expression indicates poor prognosis and promotes gastric cancer cell proliferation and invasion via upregulating Integrin/FAK/Akt pathway

The limb-bud and heart development (LBH) gene is a highly conserved, tissue-specific transcription cofactor in vertebrates that regulates multiple key genes in embryonic development. The role of LBH in various cancer types is still controversial, and its specific role and molecular mechanism in the oncogenesis of gastric cancer (GC) remains largely unexplored. In the present study, the prognostic significance and clinicopathological characteristics of LBH in GC was determined. The LBH mRNA expression was first investigated in four independent public datasets (TCGA-STAD, GSE15459, GSE29272, and GSE62254) and then validated with our samples at the protein level. LBH was overexpressed at both the mRNA and protein levels in cancer compared with normal tissues. High LBH expression was correlated with advanced T, N, and M stages. Kaplan–Meier analysis and log-rank test indicated that higher LBH expression was statistically correlated with shorter overall survival (OS) in the public datasets and our study samples. Univariate and multivariate Cox regression analysis showed that LBH was an independent prognostic biomarker for survival in TCGA-STAD, GSE15459, GSE62254 cohorts, and our GC patients. In vitro experiments showed that knockdown of LBH can significantly inhibit the proliferation and invasion of HGC-27 cells, while overexpression of LBH can significantly enhance the proliferation and invasion of BGC-823 cells. Gene Set Enrichment Analysis (GSEA), Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) indicated that high LBH expression is associated with the PI3K-Akt pathway, focal adhesion, and extracellular matrix (ECM)-receptor interaction. Western blot analysis showed that knockdown of LBH significantly inhibited the expression of integrin α5, integrin β1, p-FAK, and p-Akt. Therefore, results from the present study indicate that LBH is a potential independent prognostic biomarker and promotes proliferation and invasion of GC cells by activating the integrin/FAK/Akt pathway

Genome-Wide Association Analysis in Asthma Subjects Identifies SPATS2L as a Novel Bronchodilator Response Gene

Bronchodilator response (BDR) is an important asthma phenotype that measures reversibility of airway obstruction by comparing lung function (i.e. FEV1) before and after the administration of a short-acting β2-agonist, the most common rescue medications used for the treatment of asthma. BDR also serves as a test of β2-agonist efficacy. BDR is a complex trait that is partly under genetic control. A genome-wide association study (GWAS) of BDR, quantified as percent change in baseline FEV1 after administration of a β2-agonist, was performed with 1,644 non-Hispanic white asthmatic subjects from six drug clinical trials: CAMP, LOCCS, LODO, a medication trial conducted by Sepracor, CARE, and ACRN. Data for 469,884 single-nucleotide polymorphisms (SNPs) were used to measure the association of SNPs with BDR using a linear regression model, while adjusting for age, sex, and height. Replication of primary P-values was attempted in 501 white subjects from SARP and 550 white subjects from DAG. Experimental evidence supporting the top gene was obtained via siRNA knockdown and Western blotting analyses. The lowest overall combined P-value was 9.7E-07 for SNP rs295137, near the SPATS2L gene. Among subjects in the primary analysis, those with rs295137 TT genotype had a median BDR of 16.0 (IQR = [6.2, 32.4]), while those with CC or TC genotypes had a median BDR of 10.9 (IQR = [5.0, 22.2]). SPATS2L mRNA knockdown resulted in increased β2-adrenergic receptor levels. Our results suggest that SPATS2L may be an important regulator of β2-adrenergic receptor down-regulation and that there is promise in gaining a better understanding of the biological mechanisms of differential response to β2-agonists through GWAS

The Energy Consumption and CO2 emission impacts of Fuel and Electric Vehicles in China

Vehicle electrification has been seriously considered as an industrial revolution to achieve sustainable transportation in China. With the development of the electric system, electric vehicles and railway systems are utilizing and generalizing. To accurately and adequately evaluate the energy consumption and CO2 emission effects of fuel and electric vehicles, this study focuses on these two kinds of vehicles, including buses and cars, and analyses their environmental impacts. Results show that electric vehicles consume less heat energy and release less CO2 emission than fuel vehicles do at the same distance. Public vehicles consume much less heat energy than private cars no matter which kind of fuel they use. In addition, the CO2 emission of fuel cars is larger than that of fuel buses. Moreover, electric energy is cheaper than gasoline and diesel. Therefore, the electric power will benefit the environment and society. The power should be recommended to substitute oil in the future. Although electric vehicles still have some limitations, such as prolonging the life of batteries, developing new energy, and green energy can guarantee energy security and benefit the environment and reduce the emission of CO2

Neutrophil Extracellular Trapping Network Promotes the Pathogenesis of Neutrophil-associated Asthma through Macrophages

Asthma is a complex airway inflammatory disease that can be roughly classified into eosinophilic phenotype and non-eosinophilic phenotype. Most of the latter manifested as airway inflammation dominated by neutrophil infiltration, namely neutrophil-dominated asthma (NA). Neutrophil extracellular trapping (NETs) is a newly discovered antimicrobial mechanism of neutrophils; however, NETs can not only resist killing pathogenic microorganisms, but also promote tissue damage and autoimmune response. In the present study, we successfully established NA model in C57BL/6 mice and observed the increased formation of NETs. In NA mice, the free DNA abundance, the airway resistance, the cell numbers (total cell number, macrophage number, and neutrophil number), and inflammatory cytokine levels were significantly increased while the lung dynamic compliance was significantly reduced. After DNase I treatment, the above indexes in NA mice were all improved. In NA mice, either treatment with macrophage scavenger or IL-1β neutralizing antibody also improved the above-described indexes. In vitro, in human peripheral blood-derived neutrophils, PMA treatment significantly increased the formation of NETs. Furthermore, in macrophages differentiated from THP-1 monocytes, LPS or isolated NETs both significantly increased the levels of cytokines. In conclusion, NETs can stimulate macrophages to secrete IL-1β, which promotes neutrophils infiltration in the airway; infiltrated neutrophils, in turn, generates NETs, which can amplify the tissue damage caused by NETs and macrophages, inducing and aggravating NA