作者: Chen, Xing ; Fan, Shuai ; Zhai, Yunyun ; Wang, Jiang ; Kong, Zhengjie ; Zhang, Gaiping ; Han, Shichong ; He, Wenrui ; Du, Yongkun ; Yuan, Hang ; Hu, Man ; Zhang, Yuhang ; Wan, Bo

ABSTRACT:

The porcine reproductive and respiratory syndrome virus (PRRSV) can lead to severe reproductive problems in sows, pneumonia in weaned piglets, and increased mortality, significantly negatively impacting the economy. Post-translational changes are essential for the host-dependent replication and long-term infection of PRRSV. Uncertainty surrounds the function of the ubiquitin network in PRRSV infection. Here, we screened 10 deubiquitinating enzyme inhibitors and found that the ubiquitin-specific proteinase 1 (USP1) inhibitor ML323 significantly inhibited PRRSV replication in vitro . Importantly, we found that USP1 interacts with nonstructural protein 1β (Nsp1β) and deubiquitinates its K48 to increase protein stability, thereby improving PRRSV replication and viral titer. Among them, lysine at position 45 is essential for Nsp1β protein stability. In addition, deficiency of USP1 significantly reduced viral replication. Moreover, ML323 loses antagonism to PRRSV rSD16-K45R. This study reveals the mechanism by which PRRSV recruits the host factor USP1 to promote viral replication, providing a new target for PRRSV defense.

IMPORTANCE:

Deubiquitinating enzymes are critical factors in regulating host innate immunity. The porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 1β (Nsp1β) is essential for producing viral subgenomic mRNA and controlling the host immune system. The host inhibits PRRSV proliferation by ubiquitinating Nsp1β, and conversely, PRRSV recruits the host protein ubiquitin-specific proteinase 1 (USP1) to remove this restriction. Our results demonstrate the binding of USP1 to Nsp1β, revealing a balance of antagonism between PRRSV and the host. Our research identifies a brand-new PRRSV escape mechanism from the immune response.

2024-02-01·The FEBS journal

A genome‐wide CRISPR screen identifies USP1 as a novel regulator of the mammalian circadian clock

Article

作者: Liu, Dengfeng ; Zhang, Jing ; Hu, Ying ; Li, Jia-Da ; Lu, Renbin ; Li, Xin

The circadian clock is generated by a molecular timekeeping mechanism coordinating daily oscillations of physiology and behaviors in mammals. In the mammalian circadian clockwork, basic helix–loop–helix ARNT‐like protein 1 (BMAL1) is a core circadian component whose defects lead to circadian disruption and elicit behavioral arrhythmicity. To identify previously unknown regulators for circadian clocks, we searched for genes influencing BMAL1 protein level by using a CRISPR/Cas9‐based genome‐wide knockout library. As a result, we found that the deubiquitinase ubiquitin carboxyl‐terminal hydrolase 1 (USP1) positively affects BMAL1 protein abundance. Overexpression of wild‐type USP1, but not a deubiquitinase‐inactive mutant USP1, upregulated BMAL1 protein level, whereas genetic ablation of USP1 downregulated BMAL1 protein level in U2OS cells. Furthermore, treatment with USP1 inhibitors led to significant downregulation of BMAL1 protein in U2OS cells as well as mouse tissues. Subsequently, genetic ablation or pharmacological inhibition of USP1 resulted in reduced mRNA levels of a panel of clock genes and disrupted circadian rhythms in U2OS cells. Mechanistically, USP1 was able to de‐ubiquitinate BMAL1 and inhibit the proteasomal degradation of BMAL1. Interestingly, the expression of Usp1 was much higher than the other two deubiquitinases of BMAL1 (Usp2 and Usp9X) in the mouse heart, implying a tissue‐specific function of USP1 in the regulation of BMAL1 stability. Our work thus identifies deubiquitinase USP1 as a previously unknown regulator of the mammalian circadian clock and highlights the potential of genome‐wide CRISPR screens in the identification of regulators for the circadian clock.

2023-12-01·European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Inhibition of USP1 ameliorates hypertensive nephropathy through regulating oxidative stress and ferroptosis: A precise treatment via SJB3-019A nanodelivery

Article

作者: Shang, Jing ; Zheng, Jiaxin ; Gan, Lu ; Han, Yangwenxuan ; Li, Ying ; Hao, Fangyi ; Zhang, Yunzhu ; Zhang, Chunjian

Hypertensive nephropathy is second only to diabetes for the causation of chronic kidney disease worldwide. As the mortality and morbidity of hypertensive nephropathy keep increasing, it is important to elucidate its pathogenesis and develop new treatment strategies. In this study, an angiotensin II (Ang II)-induced renal cell system was established, and the expression of ubiquitin specific peptidase 1 (USP1) in human kidney (HK-2) cells was found to be regulated by Ang II treatment through quantitative RT-PCR and Western blot assay. The detection of glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and lipid reactive oxygen species (ROS) levels revealed that interference with USP1 reversed Ang II-induced oxidative stress and ferroptosis, which was enhanced by overexpression of USP1. Subsequently, USP1 inhibitor SJB3-019A loaded in MIL-100 and PEGTK was modified to fabricate SJB3-019A@MIL-PEGTK nanoparticles, which was confirmed to exhibit excellent alleviation of hypertension-induced oxidative stress and ferroptosis in renal cells both in vitro and in vivo. Our study identified an important pathogenesis of hypertensive nephropathy and SJB3-019A@MIL-PEGTK nanoparticle was used to develop an effective clinical treatment for hypertensive nephropathy.

项与 USP1 inhibitor(Shanghai Impact) 相关的新闻（医药）

2024-01-04

·Pharmaceutical Technology

Tango Therapeutics doses first patient with USP1 inhibitor trial

Tango entered a licensing agreement with pharma company Medivir to develop the asset in March 2020. Credit: AaronAmat via Getty Images. Tango Therapeutics has dosed the first patient in the Phase I/II clinical trial investigating ubiquitin-specific protease 1 (USP1) inhibitor TNG348. The multi-centre, open-label study (NCT06065059) will evaluate the safety, tolerability, and preliminary antitumour activity of TNG348 alone, and in combination with AstraZeneca ’s PARP inhibitor Lynparza (olaparib) in patients with BRCA1/2 mutant or HRD+ solid tumours. The study will enrol 140 subjects with an estimated completion date of June 2026, as per the ClinicalTrials.gov registry. The primary endpoints will determine the TNG348 dose as a monotherapy and in combination in the Phase I part of the trial and measure anti-tumour activity in the Phase II portion. Secondary endpoints include safety and tolerability. In March 2020, Tango entered a licensing agreement with pharma company Medivir to develop the asset. Under the contract, Medivir is entitled to multiple development and commercial milestone payments, as well as royalties on future sales of this candidate. Medivir’s CEO Jens Lindberg said: “The preclinical data generated by Tango for TNG348 is promising and dosing the first patient in a clinical study is encouraging for patients with HRD+ cancers. The efforts undertaken by Tango to develop TNG348 into a clinical-staged drug are impressive and we will continue to follow the clinical development of TNG348 with great anticipation.” USP1 inhibitors target a protein that plays a role in DNA repair processes within cells. Inhibiting USP1 can impact the repair mechanisms in cancer cells, potentially making them more vulnerable to other treatments, or hindering their ability to metastasise. Several other companies have USP1 inhibitors in the pipeline, including Roche and its KSQ-4279 for the treatment of ovarian cancer. In April 2021, Tango went public with the signing of a definitive merger agreement with the Boxer Capital-sponsored special purpose acquisition company BCTG Acquisition worth up to a total of approximately $353m.