Xiqian Jiang

All Publications

• Distinct Genetically Defined Neuronal Subtypes in the Motor Cortex Differentially Alter Their Firing Activity in the Parkinsonian State Chattre, G., Jiang, X., Wang, L., Chrapkiewicz, R., Zhang, Y., Li, J., Schnitzer, M. WILEY. 2024: S234

  View details for Web of Science ID 001365072300317

• Development of a Novel Amplifiable System to Quantify Hydrogen Peroxide in Living Cells. Journal of the American Chemical Society Wang, L., Lin, H., Yang, B., Jiang, X., Chen, J., Roy Chowdhury, S., Cheng, N., Nakata, P. A., Lonard, D. M., Wang, M. C., Wang, J. 2024

  Abstract

  Although many redox signaling molecules are present at low concentrations, typically ranging from micromolar to submicromolar levels, they often play essential roles in a wide range of biological pathways and disease mechanisms. However, accurately measuring low-abundant analytes has been a significant challenge due to the lack of sensitivity and quantitative capability of existing measurement methods. In this study, we introduced a novel chemically induced amplifiable system for quantifying low-abundance redox signaling molecules in living cells. We utilized H2O2 as a proof-of-concept analyte and developed a probe that quantifies cellular peroxide levels by combining the NanoBiT system with androgen receptor dimerization as a reporting mechanism. Our system demonstrated a highly sensitive response to cellular peroxide changes induced both endogenously and exogenously. Furthermore, the system can be adapted for the quantification of other signaling molecules if provided with suitable probing chemistry.

  View details for DOI 10.1021/jacs.4c05366

  View details for PubMedID 39079063

• Glutathione dynamics in subcellular compartments and implications for drug development. Current opinion in chemical biology Lin, H., Wang, L., Jiang, X., Wang, J. 2024; 81: 102505

  Abstract

  Glutathione (GSH) is a pivotal tripeptide antioxidant essential for maintaining cellular redox homeostasis and regulating diverse cellular processes. Subcellular compartmentalization of GSH underscores its multifaceted roles across various organelles including the cytosol, mitochondria, endoplasmic reticulum, and nucleus, each exhibiting distinct regulatory mechanisms. Perturbations in GSH dynamics contribute to pathophysiological conditions, emphasizing the clinical significance of understanding its intricate regulation. This review consolidates current knowledge on subcellular GSH dynamics, highlighting its implications in drug development, particularly in covalent drug design and antitumor strategies targeting intracellular GSH levels. Challenges and future directions in deciphering subcellular GSH dynamics are discussed, advocating for innovative methodologies to advance our comprehension and facilitate the development of precise therapeutic interventions based on GSH modulation.

  View details for DOI 10.1016/j.cbpa.2024.102505

  View details for PubMedID 39053236

• Fluorescent Probes and Mass Spectrometry-Based Methods to Quantify Thiols in Biological Systems. Antioxidants & redox signaling Wang, L., Jin, F., Jiang, X., Chen, J., Wang, M. C., Wang, J. 2021

  Abstract

  Fluorescent probes and mass spectrometry are the two most popular and complementary methods to quantify thiols in biological systems. In this review, we focus on the widely used and commercially available methods to detect and quantify thiols in living cells and the general approaches applied in mass spectrometry-based thiol quantification. We hope this review can serve as a general guide for redox biologists who are interested in thiol species.

  View details for DOI 10.1089/ars.2021.0204

  View details for PubMedID 34521263

• Glutathione Quantification in Live Cells with Real-Time Imaging and Flow Cytometry STAR PROTOCOLS Jiang, X., Chen, J., Wang, M. C., Wang, J. 2020; 1 (3): 100170

  Abstract

  Glutathione (GSH) is a highly dynamic, high abundance molecule regulating redox homeostasis in most mammalian cells. Traditional methods could not achieve quantification of glutathione in live cells with high spatial and temporal resolution. Here, we provide protocols on how to use reversible reaction-based ratiometric fluorescent probes, RealThiol (RT) and its derivatives, to quantify GSH globally or in specific organelles. The protocols are applicable to cultured or harvested cells through confocal imaging and flow cytometry. For complete details on the use and execution of this protocol, please refer to Chen et al. (2017) and Jiang et al. (2015, 2017, 2018a).

  View details for DOI 10.1016/j.xpro.2020.100170

  View details for Web of Science ID 001050085000008

  View details for PubMedID 33377064

  View details for PubMedCentralID PMC7757364

• Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation (Apri, 10.1038/s41590-020-0654-0, 2020) NATURE IMMUNOLOGY Mendiola, A. S., Ryu, J., Bardehle, S., Meyer-Franke, A., Ang, K., Wilson, C., Baeten, K. M., Hanspers, K., Merlini, M., Thomas, S., Petersen, M. A., Williams, A., Thomas, R., Rafalski, V. A., Meza-Acevedo, R., Tognatta, R., Yan, Z., Pfaff, S. J., Machado, M. R., Bedard, C., Coronado, P., Jiang, X., Wang, J., Pleiss, M. A., Green, A. J., Zamvil, S. S., Pico, A. R., Bruneau, B. G., Arkin, M. R., Akassoglou, K. 2020; 21 (9): 1135

  Abstract

  An amendment to this paper has been published and can be accessed via a link at the top of the paper.

  View details for DOI 10.1038/s41590-020-0754-x

  View details for Web of Science ID 000555034000003

  View details for PubMedID 32661365

• Sophoricoside is a selectiveLXRβantagonist with potent therapeutic effects on hepatic steatosis of mice PHYTOTHERAPY RESEARCH Zhang, Y., Li, F., Jiang, X., Jiang, X., Wang, Y., Zhang, H., Zhang, L., Fan, S., Xin, L., Yang, B., Ji, G., Huang, C. 2020; 34 (12): 3168-3179

  Abstract

  Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by the accumulation of triglycerides and associated with obesity, hyperlipidemia and insulin resistance. Currently, there is no therapy for NAFLD. Emerging evidences suggest that the inhibition of liver X receptor (LXR) activity may be a potential therapy for hepatic steatosis. Here, we identified that sophoricoside is a selective antagonist of LXRβ. Sophoricoside protected against obesity and glucose tolerance, and inhibited lipid accumulation in the liver of high-fat diet-induced obesity (DIO) mice and methionine and choline-deficient diet-induced nonalcoholic steatohepatitis mice. Furthermore, sophoricoside inhibited malondialdehyde, and increased superoxide dismutase and glutathione in the liver of the mice. In HepG2 cells, pretreatment with sophoricoside rescued GSH concentration decrease induced by H2 O2 treatment. Our data suggest that sophoricoside is a novel LXRβ selective antagonist and may improve glucose and lipid dysfunction, and attenuate lipid accumulation in the liver of DIO mice via anti-oxidant properties, which may be developed as a therapy for NAFLD.

  View details for DOI 10.1002/ptr.6747

  View details for Web of Science ID 000543507700001

  View details for PubMedID 32592532

• Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation NATURE IMMUNOLOGY Mendiola, A. S., Ryu, J., Bardehle, S., Meyer-Franke, A., Ang, K., Wilson, C., Baeten, K. M., Hanspers, K., Merlini, M., Thomas, S., Petersen, M. A., Williams, A., Thomas, R., Rafalski, V. A., Meza-Acevedo, R., Tognatta, R., Yan, Z., Pfaff, S. J., Machado, M. R., Bedard, C., Coronado, P., Jiang, X., Wang, J., Pleiss, M. A., Green, A. J., Zamvil, S. S., Pico, A. R., Bruneau, B. G., Arkin, M. R., Akassoglou, K. 2020; 21 (5): 513-+

  Abstract

  Oxidative stress is a central part of innate immune-induced neurodegeneration. However, the transcriptomic landscape of central nervous system (CNS) innate immune cells contributing to oxidative stress is unknown, and therapies to target their neurotoxic functions are not widely available. Here, we provide the oxidative stress innate immune cell atlas in neuroinflammatory disease and report the discovery of new druggable pathways. Transcriptional profiling of oxidative stress-producing CNS innate immune cells identified a core oxidative stress gene signature coupled to coagulation and glutathione-pathway genes shared between a microglia cluster and infiltrating macrophages. Tox-seq followed by a microglia high-throughput screen and oxidative stress gene network analysis identified the glutathione-regulating compound acivicin, with potent therapeutic effects that decrease oxidative stress and axonal damage in chronic and relapsing multiple sclerosis models. Thus, oxidative stress transcriptomics identified neurotoxic CNS innate immune populations and may enable discovery of selective neuroprotective strategies.

  View details for DOI 10.1038/s41590-020-0654-0

  View details for Web of Science ID 000526627600002

  View details for PubMedID 32284594

  View details for PubMedCentralID PMC7523413

• Quantitative Real-Time Imaging of Glutathione with Subcellular Resolution ANTIOXIDANTS & REDOX SIGNALING Jiang, X., Zhang, C., Chen, J., Choi, S., Zhou, Y., Zhao, M., Song, X., Chen, X., Maletic-Savatic, M., Palzkill, T., Moore, D., Wang, M. C., Wang, J. 2019; 30 (16): 1900-1910

  Abstract

  Quantitative imaging of glutathione (GSH) with high spatial and temporal resolution is essential for studying the roles of GSH in redox biology. To study the long-standing question of compartmentalization of GSH, especially its distribution between the nucleus and cytosol, an organelle-targeted quantitative probe is needed.We developed a reversible reaction-based ratiometric fluorescent probe-HaloRT-that can quantitatively measure GSH dynamics with subcellular resolution in real time. Using HaloRT, we quantitatively measured the GSH concentrations in the nucleus and cytosol of HeLa cells and primary hepatocytes under different treatment conditions and found no appreciable concentration gradients between these two organelles. Innovation and Conclusion: We developed the first reversible ratiometric GSH probe that can be universally targeted to any organelle of interest. Taking advantage of this new tool, we provided definitive evidence showing that GSH concentrations are not significantly different between the nucleus and cytosol, challenging the view of nuclear compartmentalization of GSH.

  View details for DOI 10.1089/ars.2018.7605

  View details for Web of Science ID 000464471500003

  View details for PubMedID 30358421

  View details for PubMedCentralID PMC6486671

• Cardiac-specific ablation of glutaredoxin 3 leads to cardiac hypertrophy and heart failure PHYSIOLOGICAL REPORTS Donelson, J., Wang, Q., Monroe, T. O., Jiang, X., Zhou, J., Yu, H., Mo, Q., Sun, Q., Marini, J. C., Wang, X., Nakata, P. A., Hirschi, K. D., Wang, J., Rodney, G. G., Wehrens, X., Cheng, N. 2019; 7 (8): e14071

  Abstract

  Growing evidence suggests that redox-sensitive proteins including glutaredoxins (Grxs) can protect cardiac muscle cells from oxidative stress-induced damage. Mammalian Grx3 has been shown to be critical in regulating cellular redox states. However, how Grx3 affects cardiac function by modulating reactive oxygen species (ROS) signaling remains unknown. In this study, we found that the expression of Grx3 in the heart is decreased during aging. To assess the physiological role of Grx3 in the heart, we generated mice in which Grx3 was conditionally deleted in cardiomyocytes (Grx3 conditional knockout (CKO) mice). Grx3 CKO mice were viable and grew indistinguishably from their littermates at young age. No difference in cardiac function was found comparing Grx3 CKO mice and littermate controls at this age. However, by the age of 12 months, Grx3 CKO mice exhibited left ventricular hypertrophy with a significant decrease in ejection fraction and fractional shortening along with a significant increase of ROS production in cardiomyocytes compared to controls. Deletion of Grx3 also impaired Ca2+ handling, caused enhanced sarcoplasmic reticulum (SR) calcium (Ca2+ ) leak, and decreased SR Ca2+ uptake. Furthermore, enhanced ROS production and alteration of Ca2+ handling in cardiomyocytes occurred, prior to cardiac dysfunction in young mice. Therefore, our findings demonstrate that Grx3 is an important factor in regulating cardiac hypertrophy and heart failure by modulating both cellular redox homeostasis and Ca2+ handling in the heart.

  View details for DOI 10.14814/phy2.14071

  View details for Web of Science ID 000472201500015

  View details for PubMedID 31033205

• Sulfur mustard resistant keratinocytes obtained elevated glutathione levels and other changes in the antioxidative defense mechanism Rothmiller, S., Schroeder, S., Strobelt, R., Wolf, M., Wang, J., Jiang, X., Worek, F., Steinritz, D., Thiermann, H., Schmidt, A. ELSEVIER IRELAND LTD. 2018: 51-61

  Abstract

  Sulfur mustard (SM) is a potent blistering chemical warfare agent, which was first used in 1917. Despite the Chemical Weapons Convention, a use was recently reported in Syria in 2015. This emphasizes the importance to develop countermeasures against chemical warfare agents. Despite intensive research, there is still no antidote or prophylaxis available against SM.The newly developed SM-resistant keratinocyte cell line HaCaT/SM was used to identify new target structures for drug development, particularly the adaptations in protective measures against oxidative stress. For this purpose, glutathione (GSH) and NAD(P)H levels, the effect of glutathione S-transferase (GST) inhibition as well as activation and expression of Nrf2, GST, glutamate cysteine ligase (GCL) and glutathione-disulfide reductase (GSR) as well as multi-drug resistance (MDR) proteins 1, 3 and 5 were investigated.The HaCaT/SM cells showed not only a better survival after treatment with SM or cytostatic drugs, but also hydrogen peroxide (H2O2). They exhibit more GSH even after SM treatment. Nrf2 levels were significantly lower. Inhibition of GST led to significantly decreased, activation to slightly higher IC50 values after SM treatment and a lower expression of GST was observed. The cells also expressed less GCLC and GSR. Expression of MDR1, MDR3 and MDR5 was higher under control conditions, but less stimulated by SM treatment. An increased NADP+/NADPH ratio as well as higher NAD+ levels were shown.In summary, an improved response of the resistant cell line to oxidative stress was observed. The underlying mechanisms are elevated GSH levels as well as lower expression of Nrf2 and its targets GCLC and GST as well as GSR and MDR1, MDR3 and MDR5. GST is an especially interesting target because its inhibition already induced a significant SM sensitivity. SM resistance also caused redox equivalent level differences. Taken together, these findings provide further insight into the mechanism of SM resistance and may open a window for novel therapeutic targets in SM therapy.

  View details for DOI 10.1016/j.toxlet.2017.11.024

  View details for Web of Science ID 000433259800009

  View details for PubMedID 29183814

  View details for PubMedCentralID PMC6235149

• Reply to 'Pitfalls in the quantitative imaging of glutathione in living cells' NATURE COMMUNICATIONS Jiang, X., Chen, J., Wang, J. 2018; 9: 1589

  View details for DOI 10.1038/s41467-018-04037-7

  View details for Web of Science ID 000430541900005

  View details for PubMedID 29686249

  View details for PubMedCentralID PMC5913335

• Challenges and Opportunities for Small-Molecule Fluorescent Probes in Redox Biology Applications ANTIOXIDANTS & REDOX SIGNALING Jiang, X., Wang, L., Carroll, S. L., Chen, J., Wang, M. C., Wang, J. 2018; 29 (6): 518-540

  Abstract

  The concentrations of reactive oxygen/nitrogen species (ROS/RNS) are critical to various biochemical processes. Small-molecule fluorescent probes have been widely used to detect and/or quantify ROS/RNS in many redox biology studies and serve as an important complementary to protein-based sensors with unique applications. Recent Advances: New sensing reactions have emerged in probe development, allowing more selective and quantitative detection of ROS/RNS, especially in live cells. Improvements have been made in sensing reactions, fluorophores, and bioavailability of probe molecules.In this review, we will not only summarize redox-related small-molecule fluorescent probes but also lay out the challenges of designing probes to help redox biologists independently evaluate the quality of reported small-molecule fluorescent probes, especially in the chemistry literature. We specifically highlight the advantages of reversibility in sensing reactions and its applications in ratiometric probe design for quantitative measurements in living cells. In addition, we compare the advantages and disadvantages of small-molecule probes and protein-based probes.The low physiological relevant concentrations of most ROS/RNS call for new sensing reactions with better selectivity, kinetics, and reversibility; fluorophores with high quantum yield, wide wavelength coverage, and Stokes shifts; and structural design with good aqueous solubility, membrane permeability, low protein interference, and organelle specificity. Antioxid. Redox Signal. 29, 518-540.

  View details for DOI 10.1089/ars.2017.7491

  View details for Web of Science ID 000425553600001

  View details for PubMedID 29320869

  View details for PubMedCentralID PMC6056262

• Quantitative real-time imaging of glutathione (vol 8, 16087, 2017) NATURE COMMUNICATIONS Jiang, X., Chen, J., Bajic, A., Zhang, C., Song, X., Carroll, S. L., Cai, Z., Tang, M., Xue, M., Cheng, N., Schaaf, C. P., Li, F., MacKenzie, K. R., Ferreon, A. M., Xia, F., Wang, M. C., Maletic-Savatic, M., Wang, J. 2017; 8: 16163

  Abstract

  This corrects the article DOI: 10.1038/ncomms16087.

  View details for DOI 10.1038/ncomms16163

  View details for Web of Science ID 000412134500001

  View details for PubMedID 28972204

  View details for PubMedCentralID PMC5628253

• Reversible Reaction-Based Fluorescent Probe for Real-Time Imaging of Glutathione Dynamics in Mitochondria ACS SENSORS Chen, J., Jiang, X., Zhang, C., MacKenzie, K. R., Stossi, F., Palzkill, T., Wang, M. C., Wang, J. 2017; 2 (9): 1257-1261

  Abstract

  We report a mitochondria-specific glutathione (GSH) probe-designated as Mito-RealThiol (MitoRT)-that can monitor in vivo real-time mitochondrial glutathione dynamics, and apply this probe to follow mitochondrial GSH dynamic changes in living cells for the first time. MitoRT can be utilized in confocal microscopy, super-resolution fluorescence imaging, and flow cytometry systems. Using MitoRT, we demonstrate that cells have a high priority to maintain the GSH level in mitochondria compared to the cytosol not only under normal growing conditions but also upon oxidative stress.

  View details for DOI 10.1021/acssensors.7b00425

  View details for Web of Science ID 000411853000002

  View details for PubMedID 28809477

  View details for PubMedCentralID PMC5771714

• Quantitative real-time imaging of glutathione NATURE COMMUNICATIONS Jiang, X., Chen, J., Bajic, A., Zhang, C., Song, X., Carroll, S. L., Cai, Z., Tang, M., Xue, M., Cheng, N., Schaaf, C. P., Li, F., MacKenzie, K. R., Ferreon, A. M., Xia, F., Wang, M. C., Maletic-Savatic, M., Wang, J. 2017; 8: 16087

  Abstract

  Glutathione plays many important roles in biological processes; however, the dynamic changes of glutathione concentrations in living cells remain largely unknown. Here, we report a reversible reaction-based fluorescent probe-designated as RealThiol (RT)-that can quantitatively monitor the real-time glutathione dynamics in living cells. Using RT, we observe enhanced antioxidant capability of activated neurons and dynamic glutathione changes during ferroptosis. RT is thus a versatile tool that can be used for both confocal microscopy and flow cytometry based high-throughput quantification of glutathione levels in single cells. We envision that this new glutathione probe will enable opportunities to study glutathione dynamics and transportation and expand our understanding of the physiological and pathological roles of glutathione in living cells.

  View details for DOI 10.1038/ncomms16087

  View details for Web of Science ID 000405398400001

  View details for PubMedID 28703127

  View details for PubMedCentralID PMC5511354

• Loss of glutaredoxin 3 impedes mammary lobuloalveolar development during pregnancy and lactation AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM Khanh Pham, Dong, J., Jiang, X., Qu, Y., Yu, H., Yang, Y., Olea, W., Marini, J. C., Chan, L., Wang, J., Wehrens, X. H. T., Cui, X., Li, Y., Hadsell, D. L., Cheng, N. 2017; 312 (3): E136-E149

  Abstract

  Mammalian glutaredoxin 3 (Grx3) has been shown to be important for regulating cellular redox homeostasis in the cell. Our previous studies indicate that Grx3 is significantly overexpressed in various human cancers including breast cancer and demonstrate that Grx3 controls cancer cell growth and invasion by regulating reactive oxygen species (ROS) and NF-κB signaling pathways. However, it remains to be determined whether Grx3 is required for normal mammary gland development and how it contributes to epithelial cell proliferation and differentiation in vivo. In the present study, we examined Grx3 expression in different cell types within the developing mouse mammary gland (MG) and found enhanced expression of Grx3 at pregnancy and lactation stages. To assess the physiological role of Grx3 in MG, we generated the mutant mice in which Grx3 was deleted specifically in mammary epithelial cells (MECs). Although the reduction of Grx3 expression had only minimal effects on mammary ductal development in virgin mice, it did reduce alveolar density during pregnancy and lactation. The impairment of lobuloalveolar development was associated with high levels of ROS accumulation and reduced expression of milk protein genes. In addition, proliferative gene expression was significantly suppressed with proliferation defects occurring in knockout MECs during alveolar development compared with wild-type controls. Therefore, our findings suggest that Grx3 is a key regulator of ROS in vivo and is involved in pregnancy-dependent mammary gland development and secretory activation through modulating cellular ROS.

  View details for DOI 10.1152/ajpendo.00150.2016

  View details for Web of Science ID 000395805500002

  View details for PubMedID 27894063

  View details for PubMedCentralID PMC5374299

• Genetically anchored fluorescent probes for subcellular specific imaging of hydrogen sulfide ANALYST Chen, J., Zhao, M., Jiang, X., Sizovs, A., Wang, M. C., Provost, C. R., Huang, J., Wang, J. 2016; 141 (4): 1209-1213

  Abstract

  Imaging hydrogen sulfide (H2S) at the subcellular resolution will greatly improve the understanding of functions of this signaling molecule. Taking advantage of the protein labeling technologies, we report a general strategy for the development of organelle specific H2S probes, which enables sub-cellular H2S imaging essentially in any organelles of interest.

  View details for DOI 10.1039/c5an02497h

  View details for Web of Science ID 000369617600004

  View details for PubMedID 26806071

  View details for PubMedCentralID PMC4747831