De novo design of a novel AIE fluorescent probe tailored to autophagy visualization via pH manipulation

Huang Xueyan,Chen Fei,Ma Yeshuo,Zheng Fan,Fang Yanpeng,Feng Bin,Huang Shuai,Zeng Hongliang,Zeng Wenbin 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00

Macroautophagy is an essential cellular self-protection mechanism, and defective autophagy has been considered to contribute to a variety of diseases. During the process, cytoplasmic components are transported via autophagosomes to acidic lysosomes for metabolism and recycling, which represents application niches for lysosome-targeted fluorescent probes. Additionally, in view of the complexity of the autophagy pathway, it entails more stringent requirements for probes suitable for monitoring autophagy. Meanwhile, aggregation-induced emission (AIE) fluorescent probes have been impressively demonstrated in the biomedical field, which bring fascinating possibilities to the autophagy visualization.We reported a generalizable de novo design of a novel pH-sensitive AIE probe ASMP-AP tailored to lysosome targeting for the interpretation of autophagy. Firstly, the theoretical calculation was carried out followed by the investigation of optical properties. Then, the performance of ASMP-AP in visualizing autophagy was corroborated by starvation or drugs treatments. Furthermore, the capability of ASMP-AP to monitor autophagy was demonstrated in ex vivo liver tissue and zebrafish in vivo.ASMP-AP displays a large stokes shift, great cell permeability and good biocompatibility. More importantly, ASMP-AP enables a good linear response to pH, which derives from the fact that its aggregation state can be manipulated by the acidity. It was successfully applied for imaging autophagy in living cells and was proved capable of monitoring mitophagy. Moreover, this novel molecular tool was validated by ex vivo visualization of activated autophagy in drug-induced liver injury model. Interestingly, it provided a meaningful pharmacological insight that the melanin inhibitor 1-phenyl-2-thiourea (PTU)-induced autophagy was clearly presented in wild-type zebrafish.ASMP-AP offers a simple yet effective tool for studying lysosome and autophagy. This is the first instance to visualize autophagy in zebrafish using a small-molecule probe with AIE characters, accurate lysosome targeting and simultaneous pH sensitivity. Ultimately, this novel fluorescent system has great potential for in vivo translation to fuel autophagy research.

Ratiometric and discriminative visualization of autophagic processes with a novel dual-responded lysosome-specific fluorescent probe

Fan Zheng,Yeshuo Ma,Jipeng Ding,Shuai Huang,Shengwang Zhang,Xueyan Huang,Bin Feng,Hongliang Zeng,Fei Chen,Wenbin Zeng 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00

Background Autophagy is a critical self-eating pathway involved in numerous physiological and pathological processes. Lysosomal degradation of dysfunctional organelles and invading microorganisms is central to the autophagy mechanism and essential for combating disease-related conditions. Therefore, monitoring fluctuations in the lysosomal microenvironment is vital for tracking the dynamic process of autophagy. Although much effort has been put into designing probes for measuring lysosomal viscosity or pH separately, there is a need to validate the concurrent imaging of the two elements to enhance the understanding of the dynamic progression of autophagy. Methods Probe HFI was synthesized in three steps and was developed to visualize changes in viscosity and pH within lysosomes for real-time autophagy tracking. Then, the spectrometric determination was carried out. Next, the probe was applied to image autophagy in cells under nutrient-deprivation or external stress. Additionally, the performance of HFI to monitor autophagy was employed to evaluate acetaminophen-induced liver injury. Results We constructed a ratiometric dual-responsive probe, HFI, with a large Stokes shift over 200 nm, dualwavelength emission, and small background interference. The ratiometric fluorescent signal (R = I610/I460) of HFI had an excellent correlation with both viscosity and pH. More importantly, high viscosity and low pH had a synergistic promotion effect on the emission intensity of HFI, which enabled it to specially lit lysosomes without disturbing the inherent microenvironment. We then successfully used HFI to monitor intracellular autophagy induced by starvation or drugs in real-time. Interestingly, HFI also enabled us to visualize the occurrence of autophagy in the liver tissue of a DILI model, as well as the reversible effect of hepatoprotective drugs on this event. Conclusions In this study, we developed the first ratiometric dual-responsive fluorescent probe, HFI, for real-time revealing autophagic details. It could image lysosomes with minimal perturbation to their inherent pH, allowing us to track changes in lysosomal viscosity and pH in living cells. Ultimately, HFI has great potential to serve as a useful indicator for autophagic changes in viscosity and pH in complex biological samples and can also be used to assess drug safety.

Developing a Dynamic Materialized View Index for Efficiently Discovering Usable Views for Progressive Queries

( Chao Zhu ),( Qiang Zhu ),( Calisto Zuzarte ),( Wenbin Ma ) 한국정보처리학회 2013 Journal of information processing systems Vol.9 No.4

Numerous data intensive applications demand the efficient processing of a new type of query, which is called a progressive query (PQ). A PQ consists of a set of unpredictable but inter-related step-queries (SQ) that are specified by its user in a sequence of steps. A conventional DBMS was not designed to efficiently process such PQs. In our earlier work, we introduced a materialized view based approach for efficiently processing PQs, where the focus was on selecting promising views for materialization. The problem of how to efficiently find usable views from the materialized set in order to answer the SQs for a PQ remains open. In this paper, we present a new index technique, called the Dynamic Materialized View Index (DMVI), to rapidly discover usable views for answering a given SQ. The structure of the proposed index is a special ordered tree where the SQ domain tables are used as search keys and some bitmaps are kept at the leaf nodes for refined filtering. A two-level priority rule is adopted to order domain tables in the tree, which facilitates the efficient maintenance of the tree by taking into account the dynamic characteristics of various types of materialized views for PQs. The bitmap encoding methods and the strategies/algorithms to construct, search, and maintain the DMVI are suggested. The extensive experimental results demonstrate that our index technique is quite promising in improving the performance of the materialized view based query processing approach for PQs

Developing a Dynamic Materialized View Index for Efficiently Discovering Usable Views for Progressive Queries

Zhu, Chao,Zhu, Qiang,Zuzarte, Calisto,Ma, Wenbin Korea Information Processing Society 2013 Journal of information processing systems Vol.9 No.4

Numerous data intensive applications demand the efficient processing of a new type of query, which is called a progressive query (PQ). A PQ consists of a set of unpredictable but inter-related step-queries (SQ) that are specified by its user in a sequence of steps. A conventional DBMS was not designed to efficiently process such PQs. In our earlier work, we introduced a materialized view based approach for efficiently processing PQs, where the focus was on selecting promising views for materialization. The problem of how to efficiently find usable views from the materialized set in order to answer the SQs for a PQ remains open. In this paper, we present a new index technique, called the Dynamic Materialized View Index (DMVI), to rapidly discover usable views for answering a given SQ. The structure of the proposed index is a special ordered tree where the SQ domain tables are used as search keys and some bitmaps are kept at the leaf nodes for refined filtering. A two-level priority rule is adopted to order domain tables in the tree, which facilitates the efficient maintenance of the tree by taking into account the dynamic characteristics of various types of materialized views for PQs. The bitmap encoding methods and the strategies/algorithms to construct, search, and maintain the DMVI are suggested. The extensive experimental results demonstrate that our index technique is quite promising in improving the performance of the materialized view based query processing approach for PQs.

Cytosolic escape of mitochondrial DNA triggers cGAS-STING-NLRP3 axis-dependent nucleus pulposus cell pyroptosis

Zhang Weifeng,Li Gaocai,Luo Rongjin,Lei Jie,Song Yu,Wang Bingjin,Ma Liang,Liao Zhiwei,Ke Wencan,Liu Hui,Hua Wenbin,Zhao Kangcheng,Feng Xiaobo,Wu Xinghuo,Zhang Yukun,Wang Kun,Yang Cao 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Low back pain (LBP) is a major musculoskeletal disorder and the socioeconomic problem with a high prevalence that mainly involves intervertebral disc (IVD) degeneration, characterized by progressive nucleus pulposus (NP) cell death and the development of an inflammatory microenvironment in NP tissue. Excessively accumulated cytosolic DNA acts as a damage-associated molecular pattern (DAMP) that is monitored by the cGAS-STING axis to trigger the immune response in many degenerative diseases. NLRP3 inflammasome-dependent pyroptosis is a type of inflammatory programmed death that promotes a chronic inflammatory response and tissue degeneration. However, the relationship between the cGAS-STING axis and NLRP3 inflammasome-induced pyroptosis in the pathogenesis of IVD degeneration remains unclear. Here, we used magnetic resonance imaging (MRI) and histopathology to demonstrate that cGAS, STING, and NLRP3 are associated with the degree of IVD degeneration. Oxidative stress induced cGAS-STING axis activation and NLRP3 inflammasome-mediated pyroptosis in a STING-dependent manner in human NP cells. Interestingly, the canonical morphological and functional characteristics of mitochondrial permeability transition pore (mPTP) opening with the cytosolic escape of mitochondrial DNA (mtDNA) were observed in human NP cells under oxidative stress. Furthermore, the administration of a specific pharmacological inhibitor of mPTP and self-mtDNA cytosolic leakage effectively reduced NLRP3 inflammasome-mediated pyroptotic NP cell death and microenvironmental inflammation in vitro and degenerative progression in a rat disc needle puncture model. Collectively, these data highlight the critical roles of the cGAS-STING-NLRP3 axis and pyroptosis in the progression of IVD degeneration and provide promising therapeutic approaches for discogenic LBP.