Monthly Archives: April 2022

The study, published in the journal Nature Aging, lays the ground for research that explores the possibility of translating the findings into humans.

People have traditionally thought of aging as an inevitable part of life. But since the seminal work of Cynthia Kenyon in the 90s, researchers have also become aware that aging is under genetic control.

Scientists continue to be interested in finding out whether the negative effects of aging can be reduced or reversed entirely.

Currently, 16% of the United States population is 65 years or older. By 2050 this is expected to reach 22%.

The Centers for Disease Control and Prevention (CDC) note that aging increases a persons risk of various serious chronic illnesses, such as cancer, dementia, type 2 diabetes, and heart disease.

The National Institute on Aging points out that there are various things a person can do to help reduce the effects of aging.

These include staying physically active, eating a healthy diet with lots of vegetables, fruit, and whole grains, getting a good amount of quality sleep, avoiding smoking and drinking alcohol, and regularly seeing a doctor.

In 2020 the World Health Organization (WHO) published a baseline report for the Decade of Healthy Ageing, highlighting how countries can go about ensuring health and well-being as people age.

Dr. Tedros Adhanom Ghebreyesus, WHO Director-General, says that humans now live longer than at any time in history. But adding more years to life can be a mixed blessing if it is not accompanied by adding more life to years.

The Baseline Report for the Decade of Healthy Ageing has the potential to transform the way policy-makers and multiple service providers engage with older adults. We have to work together, to foster the abilities and well-being of our older generations, who continue to give us so much.

As well as lifestyle and policy changes, scientists are also exploring whether new types of medical interventions could reduce the physiological effects of aging.

The authors behind the present study have previously found that epigenetic markers in mice could be reprogrammed using the molecules Oct4, Sox2, Klf4, and cMyc. These molecules, known as Yamanaka factors, increased the lifespan and reduced the effects of aging in mice with premature aging.

Medical News Today spoke with Prof. Juan Carlos Izpisua Belmonte, of the Gene Expression Laboratory at the Salk Institute for Biological Studies, San Diego, CA, and a corresponding author of the present study.

In the 2016 paper, we developed a protocol and showed for the first time that Yamanaka factors could be expressed in mice safely without generating cancer. Moreover, in our previous study, we used a premature aging mouse model to demonstrate that Yamanaka factors can extend the lifespan of these mice by preventing the accumulation of aging phenotypes in cells and tissues.

However, we did not know if expressing the Yamanaka factors for an extended period of time in animals without any preexisting pathologies will work and whether it would be safe. The goal of the current study was to establish whether long-term partial reprogramming would have a positive or negative impact on a wild-type animals health, said Prof. Izpisua Belmonte.

To do this, Prof. Izpisua Belmonte and his colleagues split the mice into three groups. The first group received Yamanaka factors from 15 to 22 months or around 50 to 70 years in human terms.

The second group received the Yamanaka factors from 12 to 22 months or 35 to 70 in human years.

The third group was treated for a single month at 25 months or 80 years in human terms.

The researchers found that compared with mice that acted as a control, the mice who received the Yamanaka factors did not develop cancer or see any blood cell or neurological changes.

Further, the mice that received the Yamanaka factors for a number of months showed various reversals in the effects of aging.

The kidneys and skin of the mice resembled those of younger mice, their skin healed from wounds without producing as much scarring, and the scientists did not observe the usual metabolic changes in the blood typically seen in older animals.

The animals treated for just a single month late in life did not see these effects.

Prof. Izpisua Belmonte said there were still necessary steps before the research could be tested in humans.

The translation of our approach to humans requires developing ways to deliver the factors and controlling the levels and how long the factors are expressed. These steps will allow [us] to demonstrate the safe delivery of the factors, a critical aspect before we could start thinking in clinical trials.

Nonetheless, the findings provide exciting evidence that the technique could have benefits far beyond the reversal of the effects of aging.

After our initial 2016 study, our lab, as well as several other laboratories around the world, have used the same approach to demonstrate improvement in the regeneration of different tissues in mice and rejuvenation of human cells.

All these studies further prove that the controlled expression of Yamanaka factors for cell reprogramming could benefit diverse conditions and might be a general medicine approach in the future for various complications that arise during life, said Prof. Izpisua Belmonte.

Read more from the original source:
Aging safely reversed in mice by reprogramming cells - Medical News Today

image:Intensities of protein expression of markers are shown on viSNE map as spectrum colored dots, with low in blue, high in red. view more

Credit: Bachmaier, et al. ACS Nano

Using a protein nanoparticle they designed, scientists at the University of Illinois Chicago have identified two distinct subtypes of neutrophils and found that one of the subtypes can be used as a drug target for inflammatory diseases.

Neutrophils are a type of white blood cell that help fight infection, clear dead cell debris, and heal tissue injury. But for people with health conditions caused by chronic inflammation, like arthritis or Crohns disease, or excessive inflammation, like sepsis, the role of neutrophils may be deleterious. Neutrophils have been described in research as also contributing to tissue damage the double-edged sword of inflammation. Unfortunately, current drugs for inflammatory diseases that target neutrophils suppress all their effects, including their anti-infection and healing functions.

The UIC team is the first to characterize neutrophils into two subsets.

Understanding the differences between these neutrophil subsets opens the door for more research on treatments that address inflammatory diseases without increasing patients risks of infections, said study author Kurt Bachmaier, assistant professor in the department of pharmacology and regenerative medicine at the College of Medicine, who led the research.

Bachmaier and his colleagues first used the nanoparticle platform, formulated from a protein called albumin, to analyze how neutrophils from bone marrow, blood, and spleen and lung tissues interact with the nanoparticle. They found that some neutrophils brought the albumin nanoparticle into the cell through a process called endocytosis, while others didnt.

The scientists labeled the subtype that readily endocytosed the nanoparticle as ANP-high, for albumin nanoparticle high. The neutrophils that did not absorb the albumin nanoparticle were labeled as ANP-low.

Further investigation with the albumin nanoparticle showed that the subtypes have different cell surface receptors and that they are functionally distinct in their helpful capacities to kill bacteria and their harmful potential to promote inflammation. ANP-highneutrophils did not help to kill bacteria but produced inordinate amounts of reactive oxygen species and inflammatory chemokines and cytokines, which contribute to inflammatory disease.

Because the ANP-high neutrophils are also the ones that captured the nanoparticle, the scientists conducted clever experiments using the albumin nanoparticle to deliver drug treatments. They filled the nanoparticle with an anti-inflammatory drug and administered it to mice with sepsis. They found that the mice treated with the drug-loaded nanoparticle had reduced signs of tissue inflammation, but that the neutrophilic host-defense was preserved.

The albumin nanoparticle, which was filled with the drug, specifically bound to ANP-high neutrophils and unloaded their cargo into the cell, stopping it in its tracks, Bachmaier said.We found ANP-high neutrophils not only in mice but also in humans, opening the possibility of neutrophil subset-specific targeted therapy for human inflammatory diseases.

Science can be a bit like magic by targeting only the ANP-high neutrophils, we stopped the out-of-control inflammation while preserving the bacteria-fighting inflammation of these Janus-like cells, said senior author Asrar Malik, Schweppe Family Distinguished Professor and head of the department of pharmacology and regenerative medicine.

These findings are reported in the article Albumin Nanoparticle Endocytosing Subset of Neutrophils for Precision Therapeutic Targeting of Inflammatory Tissue Injury, which is published in ACS Nano, a scientific publication of the American Chemical Society and the primary nanotechnology journal.

Co-authors of the article are Andrew Stuart, Amitabha Mukhopadhyay, Sreeparna Chakraborty, Zhigang Hong, Li Wang, Yoshikazu Tsukasaki, Mark Maienschein-Cline, Balaji Ganesh, Prasad Kanteti and Jalees Rehman.

Experimental study

Animals

Albumin Nanoparticle Endocytosing Subset of Neutrophils for Precision Therapeutic Targeting of Inflammatory Tissue Injury

1-Mar-2022

Disclaimer: AAAS and EurekAlert! are not responsible for the accuracy of news releases posted to EurekAlert! by contributing institutions or for the use of any information through the EurekAlert system.

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Newly identified neutrophil subset is a promising therapeutic target - EurekAlert

Newswise Robert Gallo, MD, The Homer & Martha Gudelsky Distinguished Professor in Medicine, co-founder and director of the Institute Human Virology at the University of Maryland School of Medicine and co-founder and Chair of the Scientific Leadership Board of the Global Virus Network, was awarded the Distinguished Alumni Award by the University of Chicago Medical & Biological Sciences Alumni Association (UChicago MBSAA) for his lifetime achievements. Honorees will participate in a panel discussion on May 10 and will be presented the award on May 21 at the Hyde Parke campus.

Being at the University of Chicago was a great inspiration for me by being so surrounded by excellence and by mentors who delighted in helping the beginning physician-scientist, said Robert C. Gallo, MD, 65.I will never forget those days which deeply impacted my entire career. Obviously, I am honored and grateful to receive this recognition.

The University of Chicago uniquely nurtured scientific discovery in Dr. Gallo's training in the practice of medicine, said I. David Goldman, MD, 62, Susan Fischer Chair, Albert Einstein College of Medicine and Director Emeritus, Albert Einstein Cancer Center.This was followed by the National Cancer Institute's recognition of the extraordinary talent and passion of a young physician-scientist providing Dr. Gallo with the freedom and resources to go on to make seminal discoveries on the biology of retroviruses and human T-cells that culminated in unraveling the causation of human T-cell leukemia and AIDS.Dr. Goldman is a former resident of the University of Chicago.

The UChicago MBSAA takes great pride in recognizing our alumni who, through their work, have made significant contributions to the biological sciences and medicine, said Mark R. Aschliman, MD'80, Chair, Alumni Awards Committee.

Dean E. Albert Reece, MD, PhD, MBA, Executive Vice President for Medical Affairs, University of Maryland Baltimore, and the John Z. and Akiko K. Bowers Distinguished Professor at the University of Maryland School of Medicine, said, My sincerest congratulations to Dr. Robert Gallo for receiving this prestigious award from one of our most preeminent academic medical institutions. Dr. Gallo is a world-renowned scientist whose breakthrough discoveries and scholarly contributions have made major contributions to global health for more than four decades. He is a visionary investigator who has unlocked many important mysteries of human viruses and diseases. He embodies all of the attributes of what it means to be a great scientist. We have been fortunate to have him as one of our most distinguished members of the University of Maryland School of Medicine faculty for many years. He has led our Institute of Human Virology, which has been transformative in its work to eradicate chronic and deadly viral and immune disorders. He is most deserving of this honor from the University of Chicago Medical Alumni Association.

Dr. Gallo graduated from Thomas Jefferson University School of Medicine before completing his medical training at the University of Chicago. After 30 years at the National Cancer Institute at the National Institutes of Health in Bethesda, Maryland, he became the co-founder of the Institute of Human Virology and the founding director and The Homer & Martha Gudelsky Distinguished Professor of Medicine and Microbiology and Immunology at the University of Maryland School of Medicine. In 2011, Dr. Gallo became the Co-Founder and Chair of the Scientific Leadership Board to the Global Virus Network.

Dr. Gallos career interests have focused on studying the basic biology of human blood cells, their normal and abnormal growth, and the involvement of viruses in these abnormalities.

Dr. Gallo and his co-workers pioneered human retrovirology, discovering the first human retrovirus (HTLV-1) and, along with others, showing it was a cause of a particular form of human leukemia. A year later, he and his group discovered the second known human retrovirus (HTLV-2). Dr. Gallo and his colleagues independently discovered HIV and provided the first results to show it was the cause of AIDS. They also developed a lifesaving HIV blood test. In 1986, he and his co-workers discovered the first new human herpes in more than 25 years, Human Herpes Virus-6 (HHV-6). Previously in 1978, Gallo discovered a variant of gibbon ape leukemia virusHalls Island strainwhich causes T-cell leukemia.

Dr. Gallo and his co-workers discovered Interleukin-2 in 1976, thus setting the stage for all groups to culture human T-cells. Gallo and his co-workers spent years developing detailed biochemical and immunological characteristics of human cellular DNA polymerases alpha, beta, and gamma and reverse transcriptase (RT) from several retroviruses to use RT as a sensitive and specific surrogate marker for retroviruses.

In 1995 he and his colleagues discovered the first natural (endogenous) inhibitors of HIV, which led to the discovery of the HIV co-receptor, CCR5, and opened new approaches to treatment. Currently, Dr. Gallo and his team have been working on a HIV preventive vaccine candidate.

Dr. Gallo has received 35 honorary doctorates from universities around the world. He was the most cited scientist from 1980 to 1990 and was ranked third in the world for scientific impact from 1983 to 2002, publishing nearly 1,300 papers.

Dr. Gallo is a member of the National Academy of Sciences and the National Academy of Medicine and has received several international prizes, including the U.S. Albert Lasker Award twice.

About the Institute of Human Virology

Formed in 1996 as a partnership between the State of Maryland, the City of Baltimore, the University System of Maryland, and the University of Maryland Medical System, the IHV is an institute of the University of Maryland School of Medicine and is home to some of the most globally-recognized and world-renowned experts in all of virology. The IHV combines the disciplines of basic research, epidemiology, and clinical research in a concerted effort to speed the discovery of diagnostics and therapeutics for a wide variety of chronic and deadly viral and immune disorders - most notably, HIV the virus that causes AIDS. For more information, visit http://www.ihv.org and follow us on Twitter @IHVmaryland.

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UM School of Medicine Institute of Human Virologys Robert Gallo Receives Distinguished Alumni Award by the University of Chicago Medical Association -...

Bo Xie,1,* Yi Chen,2,* Yebei Hu,2 Yan Zhao,2 Haixin Luo,2 Jinhui Xu,1 Xiuzu Song1

1Department of Dermatology, Hangzhou Third Peoples Hospital, Affiliated Hangzhou Dermatology Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, Peoples Republic of China; 2Department of Dermatology, Hangzhou Third Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, 310009, Peoples Republic of China

Correspondence: Xiuzu Song, Department of Dermatology, Hangzhou Third Peoples Hospital, Affiliated Hangzhou Dermatology Hospital, Zhejiang University School of Medicine, West Lake Road 38, Hangzhou, 310009, Peoples Republic of China, Tel +86-571-87823102, Email [emailprotected]

Objective: The treatment of vitiligo is often challenging to dermatologists. There is ample evidence to suggest that hydroxychloroquine (HCQ) is effective for vitiligo treatment; nonetheless, the underlying mechanism remains unknown. In the present study, we sought to uncover the molecular targets of HCQ by an integrated network-based pharmacologic and transcriptomic approach.Methods: The potential targets of HCQ were retrieved from databases based on the crystal structure. Targets related to vitiligo were screened and intersected with potential targets of HCQ. A protein-protein interaction network of the intersected targets was generated. Interactions between the targets were verified by molecular docking. Moreover, human vitiligo immortalized melanocytes (PIG3V) were evaluated after treatment with HCQ (1g/mL) for 24h. The total RNA of PIG3V was extracted and determined by RNA-seq transcriptomics for differential gene expression analysis. Network pharmacology was then used to identify the relationships between putative targets of HCQ and differentially expressed genes.Results: Molecular docking analysis revealed four putative key targets (ACHE, PNMT, MC1R, and VDR) of HCQ played important roles in vitiligo treatment. According to the transcriptomic results, the melanosomal biogenesis-related gene BLOC1S5 was upregulated 138005.020 fold after HCQ treatment. Genes related to protein repair (MSRB3) and anti-ultraviolet (UV) effect (UVSSA) were upregulated 4.253 and 2.603 fold, respectively, after HCQ treatment.Conclusion: The expression of the BLOC1S5 gene is significantly upregulated, indicating upregulated melanosomal biogenesis after HCQ treatment. In addition, HCQ yields a protective effect on melanocytes by upregulating genes associated with damaged protein repair (MSRB3) and anti-UV effect (UVSSA). The protective effects of HCQ are mediated by binding to putative targets ACHE, PNMT, MC1R, and VDR according to network pharmacology and docking verification.

Keywords: vitiligo, hydroxychloroquine, treatment, pigmentation, melanocyte protection

Vitiligo is an autoimmune depigmentation disorder characterized by the loss of functional melanocytes and patchy skin pigmentation.1 It has been reported that at least 0.5% of the population suffers from vitiligo worldwide.2 Importantly, vitiligo can lead to stigma, shame and embarrassment in this patient population.3 Until now, the mechanisms that belie the pathogenesis of vitiligo remain unknown. An increasing body of evidence suggests that both the adaptive and innate immune systems are involved in the pathogenesis of vitiligo.4 Moreover, interferon (IFN) -inducible chemokines and CD8+ T cells can be initiated by external triggers such as ultraviolet (UV) and chemical stimuli.5 Mitochondria generate reactive oxygen species (ROS) in response to oxidative stimuli, disrupting the normal functions of organelles such as mitochondria, lysosome, endoplasmic reticulum (ER), etc.6 ER injury has been reported to lead to the generation of unfolded proteins.7 Besides, it has been established that damaged proteins and exosomes secreted by melanocytes can be recognized by antigen-presenting cells to stimulate autoreactive T cell maturation.8 The positive feedback of melanocyte-specific CD8+ T cells recruitment induced by chemokines can reportedly potentiate the autoimmune attack towards melanocytes.9 Notwithstanding that unprecedented progress has been achieved in understanding the pathophysiology, the specific mechanisms have not been clearly elucidated, accounting for the difficulty dermatologists face in treating this disease during clinical practice, hence emphasizing the need for future studies.10

According to current guidelines, topical glucocorticoids, calcineurin inhibitors, vitamin D3 derivatives, and phototherapy remain the mainstay of treatment for vitiligo. Systemic glucocorticoids are indicated with rapid progression of the lesions.14 Indeed, efficient vitiligo treatment is often difficult, with low repigmentation rates and high relapse rates. Currently, HCQ is recommended to treat rheumatic diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), antiphospholipid antibody syndrome, and Sjogrens syndrome, which are autoimmune-related.1113 In addition, HCQ is reportedly effective in treating vitiligo, chronic actinic dermatitis, chronic urticaria, dermatomyositis, vasculitis, lichen planus, etc.14 In RA and SLE patients presenting with vitiligo, it was observed that HCQ treatment also promotes pigmentation in vitiligo lesions.10,15 In addition, skin pigmentation after HCQ treatment was observed in RA and SLE patients without vitiligo.16,17 It is widely believed that HCQ exerts an anti-inflammatory effect that can disrupt T-cell receptor-related Ca2+ signaling and antigen processing.18 Importantly, Li DG et al19 showed that HCQ could protect melanocytes from autoantibody-induced damage by reducing the formation of antigen-antibody complexes.

At present, the mechanisms underlying the therapeutic effect of HCQ in vitiligo are unclear. Current evidence suggests that HCQ could be a multi-target drug given its wide range of effects. Indeed, the immunoregulatory effect of HCQ plays a key role in its pharmacological mechanisms. In recent years, the rapid development of computer technology and big data analytics has led to the advent of network pharmacology, which provides a new strategy for the research of multi-targets drugs. Network pharmacology integrates poly-pharmacology, bioinformatics and systems biology for multi-targets drug research and evaluation. Importantly, network pharmacology emphasizes the multi-targets network/drug pattern in contrast to the conventional one target/ one drug paradigm.2023 In the present study, human vitiligo immortalized melanocytes (PIG3V) were used to conduct differential gene expression analysis by RNA-sequencing after HCQ treatment. Transcriptomic, GO annotation, and KEGG enrichment analyses were conducted to elaborate changes in gene expression and biological functions after HCQ treatment. Finally, network pharmacology and transcriptomic results were integrated to uncover the mechanisms of HCQ in treating vitiligo, providing a foothold for future studies on its potential use for vitiligo treatment.

The predicted target proteins were retrieved from the database according to the crystal structure and chemical groups of HCQ. Then, vitiligo-related proteins were collected from disease databases. Data on these proteins were imported into Cytoscape software (v.3.8.2). The two groups of proteins were then intersected. A protein-protein interaction (PPI) network of the intersected proteins was constructed with the plugin Bisogenet of Cytoscape software. Subsequently, all intersected targets and hub proteins in the PPI network were chosen for molecular docking with HCQ. Then, gene expression in PIG3V was explored after treatment with HCQ. Transcriptomic, GO annotation and KEGG enrichment analyses were conducted. Finally, we integrated network pharmacology and transcriptomic results to assess the relationships between HCQ molecular docking targets and differentially expressed genes after HCQ treatment in PIG3V cells (Figure 1). According to the guidelines of Ethics Committee of Hangzhou Third Peoples Hospital, the human public databases we used in this research were exempted from approval, because only studies of identifiable human specimens or data need to be approved by ethics committee.

Figure 1 The flow chat of strategy layout. The mechanisms of HCQ in treating vitiligo were predicted by network pharmacology. Hub targets were verified by molecular docking. The efficacy of HCQ on vitiligo were then observed on PIG3V cell line. Differential gene expression was analyzed and linked to HCQ targets that verified by docking.

The chemical structure (Canonical SMILES) of HCQ was retrieved from PubChem database (https://pubchem.ncbi.nlm.nih.gov/): CCN(CCCC(C)NC1=C2C=CC(=CC2=NC=C1)Cl)CCO. The SMILES molecular formula of HCQ was imported into the SwissTargetPrediction database (http://www.swisstargetprediction.ch/index.php). Then predicted targets of HCQ were calculated and retrieved from the SwissTargetPrediction database based on the crystal structure. The species was set as Homo sapiens. The top 100 potential targets were selected according to the calculated parameters.24,25 Vitiligo-related proteins were searched using the keywords and species (vitiligo and Homo sapiens) across four databases, including Genecards (https://www.genecards.org/), OMIM (https://omim.org/), Drugbank (https://go.drugbank.com/) and DisGeNET (https://www.disgenet.org/). A total of 1185 vitiligo-related proteins were screened.26,27 The bisogenet plugin of Cytoscape was used for the PPI network construction. The intersected proteins between vitiligo and HCQ were entered into bisogenet. The topological features degree, betweenness and closeness were used to select the putative targets using the Cytoscape plugin CytoNCA.28

Along with the intersection targets between HCQ and vitiligo, the core targets screened in the previous step were chosen as candidates for molecular docking. The spatial interactions between target proteins and HCQ were analyzed by AutoDock (v.4.2) software. First, the 3-dimensional crystal structure of HCQ was retrieved from the Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB) database. Then, modifications such as water and ligand removal, amino acid optimization and patching, and hydrogen addition were manipulated in AutoDock. ChemBioDraw 3D (v.15.1) software was used for 3-D visualization and energy minimizing. Finally, MolegroVirtualDocker software was used to compute docking targets by comparing the conformation with the existing 3-D crystal structure of HCQ.29

The Human vitiligo melanocyte cell line PIG3V was a gift from Professor Chunying Li (Xijing Hospital of Air Force Medical University, Xian, China) and cultured (2105 cells/mL) in a 6-well plate (2mL/well) supplemented with DMEM/F12, 10% FBS, basic fibroblast growth factor (10ng/mL), phorbol myristate acetate (10ng/mL), and penicillin/streptomycin (10000U/mL, 10000g/mL) at 37C in 5% CO2 for 24 hours.30 The cultured PIG3V cells were randomly divided into an HCQ group and a control group. Cells in the HCQ group were treated with 1g/mL HCQ (Sigma-Aldrich Corp., St. Louis, MO, USA) for 24 hours. The HCQ powder was dissolved in phosphate-buffered saline (PBS). In contrast, cells in the control group were treated with PBS for 24 hours. Finally, the total RNA of PIG3V cells was purified and extracted with TRIzol reagent (Invitrogen, Carlsbad, CA, USA).

RNA samples extracted from PIG3V cells were quantified by NanoDrop (Wilmington, DE, USA) and Bioanalyzer (Agilent, CA, USA).31,32 Poly (A) RNA was extracted from total RNA by 2 rounds of purification with Dynabeads Oligo (Thermo Fisher, CA, USA). Small pieces of Poly A RNA were returned through Magnesium RNA Fragmentation Module (NEB, USA) under the condition of 94C for 5 minutes. Subsequently, SuperScript Reverse Transcriptase (Invitrogen, USA) was used to reverse-transcribe the cleaved RNA pieces into cDNA. Then, U-labeled second-stranded DNAs were synthesized. After addition of A-base to each strand, index read preparation, and heat-labile UDG enzyme treatment, PCR amplification was conducted. PCR consisted of preheating at 95C for 3 minutes; 8 cycles of 98C for 15 seconds, annealing at 60C for 15 seconds and extension at 72C for 5 minutes. Ultimately, RNA sequencing was performed according to the protocol of Illumina Novaseq 6000 (LC-Bio Technology CO., Ltd., Hangzhou, China).33,34 Genes differential expression analysis was performed by DESeq2 software between two different groups. After calculation, differentially expressed genes were screened by fold change (FC)>2 or FC<0.5 and p value<0.05. These genes underwent GO and KEGG functional enrichment analyses. Hypergeometric test was used to calculate p value in GO and KEGG enrichment.35,36

According to the value of | log2(FC) |, the top 30 upregulated and downregulated genes in PIG3V cells were selected. To screen genes related to synthesis, transport, metabolism of melanin, melanocyte protection and other mechanisms associated with vitiligo, the functions of these genes/proteins and previous studies were retrieved from the databases of UniProt (https://www.uniprot.org/) and NCBI PubMed (https://www.ncbi.nlm.nih.gov/). Then these differentially expressed genes along with docking HCQ targets were input into Cytoscape software. A PPI network was constructed to explore the interactions between HCQ targets and vitiligo-related differentially expressed genes.37,38

The predicted targets of HCQ were computed and retrieved from databases by analyzing the 2 and 3-dimensional chemical structure (Figure 2A). The top 100 potential targets (Figure 2B) were retrieved according to the index of possibility. These predicted targets were mainly classified as G protein-coupled receptors (29%), kinases (26%) and surface antigens (13%) (Figure 2C).

Figure 2 Predicted targets of HCQ. (A) 3 dimensional and 2 dimensional structure of HCQ; (B) top 100 predicted targets of HCQ from databases; (C) categories of the predicted targets.

The intersection between vitiligo-related proteins (n=1185) and top 100 potential HCQ targets yielded 15 proteins (Table 1). A PPI network of the intersected targets was generated by Cytoscape using the plugin Bisogenet. Then, 1686 additional proteins closely linked to the 15 intersected proteins were retrieved from Bisogenet. Finally, a total of 42,371 interactions (edges) between 1686 targets (nodes) were identified (Figure 3A). A topological degree greater than 62 was used to screen vital nodes in these 1686 targets (Figure 3B). Then, 459 nodes and 18,251 edges were screened according to a betweenness value greater than 734.84 and closeness value greater than 0.50. The 78 hub proteins and their 1452 interactions which may play important roles in the therapeutic effects of HCQ in treating vitiligo, were retrieved (Figure 3C). The top 10 hub proteins are listed in Table 2.

Table 1 The Intersection of HCQ Predicted Targets and Vitiligo-Related Proteins

Table 2 The Hub Proteins Screened in PPI Network

Figure 3 The PPI network conducted with 15 mutual genes of vitiligo and HCQ potential targets. (A) PPI network of the enlarged 1686 nodes; (B) the nodes and edges after the first screening; (C) the nodes and edges after the second screening.

The intersected 15 proteins and the top 10 hub proteins of the PPI network were chosen as target candidates for molecular docking verification (Tables 1 and 2), providing them visual explanations of spatial interactions with HCQ. A docking score below 20 demonstrated that HCQ could effectively combine with target proteins. Acetylcholinesterase (ACHE), Phenylethanolamine N-methyltransferase (PNMT), Melanocortin receptor 1 (MC1R), and Vitamin D3 receptor (VDR) were identified by molecular docking analysis as the putative targets of HCQ during vitiligo treatment (Table 3). The 4 key targets exhibited the most solid chemical binding forces and spatial conjunctions with HCQ. Importantly, the docking results showed that ionic bonds, hydrogen bonds and - stacking interactions were the predominant chemical forces. For instance, the hydroxyl, carbonyl and amino groups within HCQ formed hydrogen bonds with target proteins. The benzene and aromatic rings of HCQ formed - stacking interactions with target proteins (Figure 4).

Table 3 The Docking Results of Target Candidates

Figure 4 Molecular docking verification. Target candidates (A) ACHE, (B) VDR, (C) MC1R, and (D) PNMT were shown interacting with HCQ molecule (represented by a green ball-and-stick model).

PIG3V is an immortalized cell line derived from human vitiligo melanocytes. To better understand the state of PIG3V cells after HCQ (1g/L) treatment for 24 hours, we analyzed the transcriptome of PIG3V. We found 108 and 97 DEGs were upregulated and downregulated by 2 fold or more in PIG3V cells, respectively, after HCQ treatment compared to the PIG3V control (p value<0.05). The top 30 upregulated and down-regulated genes in PIG3V cells were selected according to the value of | log2(FC) | and displayed in Table 4. As for the melanin synthesis pathway, BLOC1S5 was highly upregulated by 138005.020 fold after HCQ treatment compared to the control group, suggesting that melanin synthesis was significantly enhanced. In addition, two upregulated DEGs, MSRB3 and UVSSA, were enriched in melanocyte protection, which indicated an antioxidant effect in PIG3V cells after HCQ treatment. In addition, many upregulated genes were involved in modulating the activity of the immune system during acute-phase reactions such as ORM1, SAA1, SAA2, HP, FGA, FGB, FGG, CRP, AMBP, PTPRC, SERPINA1, and SERPINA3. These acute-phase genes may be related to the antibiotic effect of HCQ, acting as guards against the invasion of harmful microorganisms such as plasmodium.39 In contrast to acute-phase genes, some chronic-phase genes were downregulated, including COL1A1, DCN, and ELN. The downregulated genes were associated with decreased collagen synthesis and fibrosis, which are features of chronic inflammation. This finding may be a potential mechanism of HCQ in treating RA and other rheumatic diseases. Moreover, current evidence suggests that COL1A1 and ELN downregulation are associated with skin aging.40,41 KEGG pathway analysis showed that platelet activation, protein digestion, absorption, and herpes simplex virus 1 infection were significantly enriched pathways. GO enrichment analysis showed that GO terms, including acute-phase response, collagen-containing extracellular matrix, and extracellular matrix organization, were significantly enriched for molecular functions (Figure 5). However, it remains unclear whether these acute-phase genes are linked to vitiligo.

Table 4 Differential Gene Expression of PIG3V After HCQ Treatment

Figure 5 Transcriptomics analysis of PIG3V cells treated by HCQ. (A) number of up-regulated and down-regulated genes; (B) heat map of up-regulated and down- regulated genes; (C) KEGG enrichment of differential expressed genes; (D) volcano map of up-regulated and down-regulated genes; (E) GO enrichment of differential expressed genes.

Abbreviations: HCQ, hydroxychloroquine treated PIG3V cells group; NG, normal control group (PIG3V without hydroxychloroquine treatment).

To validate that ACHE, PNMT, MC1R, and VDR were targets of HCQ during vitiligo treatment, the differentially expressed genes in PIG3V cells were detected by RNA sequencing. Except for acute phase genes and other genes detected by RNA sequencing, genes potentially related to vitiligo or melanocyte (including BLOC1S5, MSRB3 and UVSSA) underwent PPI network analysis with docked HCQ targets, yielding a total of 163 nodes (proteins) along with 1019 edges (interactions) (Figure 6).

Figure 6 The network between HCQ targets and differential expressed genes of PIG3V cells after the treatment of HCQ. (A) PPI network between docked HCQ targets and differential expressed genes of PIG3V showed enlarged 163 nodes (proteins) and 1019 edges (interactions); (B) main connections of the PPI network.

In the present study, the transcriptomic results showed that BLOC1S5 gene expression exhibited the highest fold change (138005.020 fold) after PIG3V cells were treated with HCQ. It has been established that BLOC1S5 encodes a subunit of the biogenesis of the lysosome-related organelles complex (BLOC-1), which is involved in melanosomal biogenesis.42 Oculocutaneous depigmentation is widely acknowledged as one of the characteristics of Hermansky-Pudlak Syndrome (HPS). Recent studies have demonstrated the presence of pathogenic BLOC1S5 variants in HPS.43,44 Moreover, it has been shown that knockdown of BLOC1S5 in zebrafish led to retinal depigmentation.45 In addition, a significant association between single nucleotide polymorphism (SNP) of 3 genes, including BLOC1S5, involved in skin pigmentation and 25-(OH)D serum concentration has been found.46 In the present study, network pharmacology analysis demonstrated that Vitamin D3 receptor (VDR) was a predicted target of HCQ, which may play a role similar to 25-(OH)D. In addition, another docking target of HCQ, MC1R, is a documented G-protein-coupled receptor that plays a vital role in skin pigmentation. It has been reported that melanocyte-stimulating hormone (-MSH) could stimulate cAMP signaling and melanin production after combination with MC1R, enhancing the repair of damaged DNA and protein after UV exposure.45 The MC1R/cAMP/MITF signaling pathway is well-established to regulate UV-induced pigmentation.46 Although no direct relationship between MC1R/VDR and BLOC1S5 was found in our PPI network, the highly upregulated BLOC1S5 indicated modulation of this pathway (Figure 7). Further studies are warranted in the future to validate this finding.

Figure 7 Illustration of related genes involved in pigmentation pathway. BLOC1S5, PMEL, DTNBP1 and PLDN are genes involved in genesis of the melanosome. MC1R, MITF, DKK1, RAB27A, MLPH and MYO5A encode proteins in membrane or cytoplasm which are involved in signaling pathways of skin pigmentation. PAX3 and SOX10 are transcription factors in melanocyte. RACK1 is a regulator to the signal transduction in melanocyte. MSRB3 and UVSSA encode proteins that are involved in damaged proteins/DNA repair under UV. MC1R, VDR, PNMT, and ACHE was targeted by HCQ according to network pharmacology and molecular docking, which could further up-regulate gene expression of BLOC1S5, MSRB3, and UVSSA, showing the promotion effect of melanosome genesis and melanocyte protection under UV damage.

Abbreviations: MSH, melanocyte stimulating hormone; MC1R, melanocortin 1 receptor; BLOC1S5, biogenesis of lysosomal organelles complex-1 subunit 5; cAMP, 3,5-cyclic adenosine monophosphate; PMEL, premelanosome protein; DTNBP1, dystrobrevin binding protein 1; PLDN, biogenesis of lysosomal organelles complex-subunit 6; MITF, microphthalmia-associated transcription factor; PAX3, paired box 3; RACK1, guanine nucleotide binding protein; SOX10, SRY (sex determining region Y)-box 10; RAB27A, member RAS oncogene family; MLPH, melanophilin; MYO5A, Myosin VA (heavy chain 12, myoxin); UV, ultraviolet; PNMT, Phenylethanolamine N-methyltransferase; ACHE, Acetylcholinesterase; VDR, Vitamin D3 receptor; Ach, acetylcholine; AD, adrenaline; NA, noradrenaline; HCQ, hydroxychloroquine.

Current evidence suggests that in vitiligo patients, organelle functions of melanocytes are damaged in response to external triggers such as UV and chemical stimuli, leading to the attack of the bodys immune system.7 Our transcriptomic analysis showed that DEGs, including MSRB3 and UVSSA, were upregulated 2 fold or more after treatment with HCQ. These two genes are widely acknowledged for protein and DNA repair and maintaining organelle homeostasis. It has been shown that MSRB3 encodes methionine-R-sulfoxide reductase (MSR). In response to environmental stress, many reactive oxygen species (ROS) are produced. ROS can oxidize methionine (Met) residues in protein peptides to form methionine sulfoxide [Met(O)], which leads to protein function impairment. Importantly, MSR could catalyze the reduction of Met(O) to restore the function of proteins in response to oxidative damage. Accordingly, MSRB3 plays a pivotal role in cell protection under environmental stress.47 Moreover, it has been shown that UVSSA encodes for the UV-stimulated scaffold protein A, a transcription-coupled nucleotide excision repair (TCNER) factor, in response to UV damage. TCNER stabilizes gene ERCC6 by recruiting the enzyme USP7 into the TCNER complex, preventing UV-induced ERCC6 degradation by proteasomes.48 TCNER induces the removal of RNA polymerase II (RNA pol II) from the active genes for transcription. Subsequently, ubiquitination at UV-damaged sites can accelerate RNA pol II recalling to nucleotide excision repair machinery.49 According to the molecular docking and PPI network analysis results (Figure 6), the MC1R-MSRB3/UVSSA axis could be an essential mechanism of HCQ in the process of melanocyte protection against oxidative stress, and the MC1R agonist could serve as a cutaneous pigmentation promoter. An increasing body of evidence suggests that activation of MC1R alleviates oxidative stress and neuronal apoptosis through protein kinase R-like endoplasmic reticulum kinase (PERK)-nuclear factor erythroid 2-related factor 2 (NRF2) pathway, exhibiting an antioxidant role in the unfolded protein response (UPR) system. As the UPR system is initiated, impaired (unfolded, misfolded, etc.) proteins are restored by enzymes through a series of reactions.7,50

As molecular docking targets of HCQ, ACHE and PNMT potentially play important roles in vitiligo treatment. Non-neuronal acetylcholine (ACh) is well-recognized to regulate human keratinocyte (KCs) functions, including cell differentiation, cell-cell interaction, secretion, and mitosis. Besides, ACh plays an important role in immune regulation. It has been established that the Ach receptor is expressed in both KCs and melanocytes.51 A study reported that the average levels of ACh were higher in areas of skin depigmentation compared to controls. Interestingly, the level of ACh was significantly decreased after treatment, which showed a significant positive correlation with the severity of vitiligo.52 A study by Taieb et al53 substantiated the toxic effect of ACh on melanocytes. In addition to ACHE, PNMT, which catalyzes the synthesis of adrenaline (AD) from noradrenaline (NA), plays an important role in vitiligo. It has been shown that KCs could synthesize NA and AD. Current evidence suggests that KCs in the lesion area of vitiligo patients synthesized 4 times more NA than normal skin area, with low PNMT activity.54 Therefore, increasing the activity of ACHE and PNMT to reduce skin ACh and NA may be one of the mechanisms underlying the efficacy of HCQ in vitiligo treatment. Nonetheless, further studies are required to increase the robustness of our findings.

This present study uncovered HCQ targets (ACHE, PNMT, MC1R, and VDR) during vitiligo treatment after screening by network pharmacology and molecular docking. PIG3V cells were used to explore the mechanisms of HCQ treatment through transcriptomic analysis. The results of transcriptomic study further showed that through the above targets, HCQ significantly promoted the expression of genes related to melanin synthesis. In addition, the expression of the genes which are related to DNA and protein damage repair was also significantly up-regulated, indicating the protective effect of HCQ on vitiligo melanocytes. In silico methods were used to identify the relationships between HCQ targets and differentially expressed genes in PIG3V cells. These findings provided the theoretical basis for the mechanisms of HCQ in treating vitiligo (Figure 7). Nevertheless, there were some limitations in our study. The specific mechanisms and signal pathways of the MC1R/VDR-BLOC1S5 axis and MC1R-MSRB3/UVSSA axis involved in the pigmentation process and melanocytes protection effect were not explored. In addition, the effect of HCQ on ACHE and PNMT and further pathways were not assessed, warranting further studies.

This work was supported by [National Natural Science Foundation of China] under Grant [number 81872517].

The authors declare that they have no conflicts of interest.

1. Ezzedine K, Eleftheriadou V, Whitton M, et al. Vitiligo. Lancet. 2015;386:7484. doi:10.1016/S0140-6736(14)60763-7

2. Bergqvist C, Ezzedine K. Vitiligo: a review. Dermatology. 2020;236:571592. doi:10.1159/000506103

3. Alikhan A, Felsten LM, Daly M, et al. Vitiligo: a comprehensive overview part I. Introduction, epidemiology, quality of life, diagnosis, differential diagnosis, associations, histopathology, etiology, and work-up. J Am Acad Dermatol. 2011;65:473491. doi:10.1016/j.jaad.2010.11.061

4. Picardo M, DellAnna ML, Ezzedine K, et al. Vitiligo. Nat Rev Dis Primers. 2015;1:15011. doi:10.1038/nrdp.2015.11

5. Strassner JP, Harris JE. Understanding mechanisms of autoimmunity through translational research in vitiligo. Curr Opin Immunol. 2016;43:8188. doi:10.1016/j.coi.2016.09.008

6. Laddha NC, Dwivedi M, Mansuri MS, et al. Role of oxidative stress and autoimmunity in onset and progression of vitiligo. Exp Dermatol. 2014;23:352353. doi:10.1111/exd.12372

7. Xie B, Song XZ. The impaired unfolded protein-premelanosome protein and transient receptor potential channels-autophagy axes in apoptotic melanocytes in vitiligo. Pigment Cell Melanoma Res. 2022;35:617. doi:10.1111/pcmr.13006

8. Al-Shobaili HA, Rasheed Z. Mitochondrial DNA acquires immunogenicity on exposure to nitrosative stress in patients with vitiligo. Hum Immunol. 2014;75:10531061. doi:10.1016/j.humimm.2014.09.003

9. Xie H, Zhou F, Liu L, et al. Vitiligo: how do oxidative stress-induced autoantigens trigger autoimmunity? J Dermatol Sci. 2016;81:39. doi:10.1016/j.jdermsci.2015.09.003

10. Joo K, Park W, Kwon SR, et al. Improvement of vitiligo in a patient with rheumatoid arthritis after hydroxychloroquine treatment. Int J Rheum Dis. 2015;18:679680. doi:10.1111/1756-185X.12442

11. Fanouriakis A, Kostopoulou M, Alunno A, et al. 2019 update of the EULAR recommendations for the management of systemic lupus erythematosus. Ann Rheum Dis. 2019;78(6):736745. doi:10.1136/annrheumdis-2019-215089

12. Smolen JS, Landew RBM, Bijlsma JWJ, et al. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2019 update. Ann Rheum Dis. 2020;79(6):685699. doi:10.1136/annrheumdis-2019-216655

13. Ramos-Casals M, Brito-Zern P, Bombardieri S, et al. EULAR recommendations for the management of Sjgrens syndrome with topical and systemic therapies. Ann Rheum Dis. 2020;79:318. doi:10.1136/annrheumdis-2019-216114

14. Chew CY, Mar A, Nikpour M, et al. Hydroxychloroquine in dermatology: new perspectives on an old drug. Australas J Dermatol. 2020;61:e150e157. doi:10.1111/ajd.13168

15. Niebel D, Braegelmann C, Bieber T, et al. Vitiligo-like depigmentation subsequent to subacute cutaneous lupus erythematosus and hydroxychloroquine treatment. J Dtsch Dermatol Ges. 2020;18:14701473.

16. Walsh DS, Farley MF, Beard JS, et al. Systemic lupus erythematosus: nephritis, dilated cardiomyopathy, and extensive cutaneous depigmentation responsive to hydroxychloroquine. J Am Acad Dermatol. 1995;33:828830. doi:10.1016/0190-9622(95)91842-6

17. Kwak D, Grimes Pearl E. A case of hyperpigmentation induced by hydroxychloroquine and quinacrine in a patient with systemic lupus erythematosus and review of the literature. Int J Womens Dermatol. 2020;6:268271. doi:10.1016/j.ijwd.2020.06.009

18. Khan DA. Alternative agents in refractory chronic urticaria: evidence and considerations on their selection and use. J Allergy Clin Immunol Pract. 2013;1(5):433440. doi:10.1016/j.jaip.2013.06.003

19. Li DG, Hu WZ, Ma HJ, et al. Hydroxychloroquine protects melanocytes from autoantibody-induced injury by reducing the binding of antigen-antibody complexes. Mol Med Rep. 2016;14:12751282. doi:10.3892/mmr.2016.5354

20. Ma C, Xu T, Sun X, et al. Network pharmacology and bioinformatics approach reveals the therapeutic mechanism of action of baicalein in hepatocellular carcinoma. Evid Based Complement Alternat Med. 2019:7518374. doi:10.1155/2019/7518374

21. Luo TT, Lu Y, Yan SK, et al. Network pharmacology in research of Chinese medicine formula: methodology, application and prospective. Chin J Integr Med. 2020;26(1):19. doi:10.1007/s11655-019-3064-0

22. Ding M, Ma W, Wang X, et al. A network pharmacology integrated pharmacokinetics strategy for uncovering pharmacological mechanism of compounds absorbed into the blood of Dan-Lou tablet on coronary heart disease. J Ethnopharmacol. 2019;242:112055. doi:10.1016/j.jep.2019.112055

23. Zhang H, Yan ZY, Wang YX, et al. Network pharmacology-based screening of the active ingredients and potential targets of the genus of Pithecellobium marthae (Britton & Killip) Niezgoda & Nevl for application to Alzheimers disease. Nat Prod Res. 2019;33(16):23682371. doi:10.1080/14786419.2018.1440222

24. Otasek D, Morris JH, Bouas J, et al. Cytoscape automation: empowering workflow-based network analysis. Genome Biol. 2019;20(1):185200. doi:10.1186/s13059-019-1758-4

25. Xie B, Geng Q, Xu J, et al. The multi-targets mechanism of hydroxychloroquine in the treatment of systemic lupus erythematosus based on network pharmacology. Lupus. 2020;29:17041711. doi:10.1177/0961203320952541

26. Stelzer G, Rosen R, Plaschkes I, et al. The genecards suite: from gene data mining to disease genome sequence analysis. Curr Protoc Bioinform. 2016;54. doi:10.1002/cpbi.5

27. Xie B, Lu H, Jinhui X, et al. Targets of hydroxychloroquine in the treatment of rheumatoid arthritis. A network pharmacology study. Joint Bone Spine. 2021;88:105099. doi:10.1016/j.jbspin.2020.105099

28. Martin A, Ochagavia ME, Rabasa LC, et al. BisoGenet: a new tool for gene network building, visualization and analysis. BMC Bioinform. 2010;11(1):9199. doi:10.1186/1471-2105-11-91

29. Rizvi SM, Shakil S, Haneef M. A simple click by click protocol to perform docking: autoDock 4.2 made easy for non-bioinformaticians. EXCLI J. 2013;12:831857.

30. Tian J, Wang Y, Ding M, et al. The formation of melanocyte apoptotic bodies in vitiligo and the relocation of vitiligo autoantigens under oxidative stress. Oxid Med Cell Longev. 2021;2021:7617839. doi:10.1155/2021/7617839

31. Beekman R, Chapaprieta V, Russiol N, et al. The reference epigenome and regulatory chromatin landscape of chronic lymphocytic leukemia. Nat Med. 2018;24(6):868880. doi:10.1038/s41591-018-0028-4

32. Wang LG, Wang SQ, Li W. RSeQC: quality control of RNA-seq experiments. Bioinformatics. 2012;28:21842185. doi:10.1093/bioinformatics/bts356

33. Wang K, Li M, Hakonarson H. ANNOVAR: functional annotation of genetic variants from next-generation sequencing data. Nucleic Acids Res. 2010;38:e164. doi:10.1093/nar/gkq603

34. Florea L, Song L, Salzberg SL. Thousands of exon skipping events differentiate among splicing patterns in sixteen human tissues. F1000Res. 2013;2:188. doi:10.12688/f1000research.2-188.v1

35. Robinson MD, McCarthy DJ, Smyth GK. edgeR: a bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26:139140. doi:10.1093/bioinformatics/btp616

36. Pertea M, Pertea GM, Antonescu CM, et al. StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol. 2015;33:290295. doi:10.1038/nbt.3122

37. Li AP, Liu Y, Cui T, et al. Uncovering the mechanism of Astragali Radix against nephrotic syndrome by intergrating lipidomics and network pharmacology. Phytomedicine. 2020;77:153274. doi:10.1016/j.phymed.2020.153274

38. Li T, Zhang W, Hu E, et al. Integrated metabolomics and network pharmacology to reveal the mechanisms of hydroxysafflor yellow A against acute traumatic brain injury. Comput Struct Biotechnol J. 2021;19:10021013. doi:10.1016/j.csbj.2021.01.033

39. Raju MVJ, Kamaraju RS, Sritharan V, et al. Continuous evaluation of changes in the serum proteome from early to late stages of sepsis caused by Klebsiella pneumoniae. Mol Med Rep. 2016;13:48354844. doi:10.3892/mmr.2016.5112

40. Bigot N, Beauchef G, Hervieu M, et al. NF-B accumulation associated with COL1A1 transactivators defects during chronological aging represses type I collagen expression through a 112/-61-bp region of the COL1A1 promoter in human skin fibroblasts. J Invest Dermatol. 2012;132:23602367. doi:10.1038/jid.2012.164

41. Osborne R, Carver RS, Mullins LA, et al. Practical application of cellular bioenergetics to the care of aged skin. Br J Dermatol. 2013;169:3238. doi:10.1111/bjd.12439

42. Rossberg W, Saternus R, Wagenpfeil S, et al. Human pigmentation, cutaneous vitamin D synthesis and evolution: variants of genes (SNPs) involved in skin pigmentation are associated with 25(OH)D serum concentration. Anticancer Res. 2016;36:14291437.

43. Pennamen P, Le L, Tingaud-Sequeira A, et al. BLOC1S5 pathogenic variants cause a new type of Hermansky-Pudlak syndrome. Genet Med. 2020;22:16131622. doi:10.1038/s41436-020-0867-5

44. Zhong Z, Wu Z, Zhang J, et al. A novel BLOC1S5-related HPS-11 patient and zebrafish with bloc1s5 disruption. Pigment Cell Melanoma Res. 2021;34:11121119. doi:10.1111/pcmr.12995

45. Rachmin I, Ostrowski SM, Weng QY, et al. Topical treatment strategies to manipulate human skin pigmentation. Adv Drug Deliv Rev. 2020;153:6571. doi:10.1016/j.addr.2020.02.002

46. Fu S, Luo X, Wu X, et al. Activation of the melanocortin-1 receptor by NDP-MSH attenuates oxidative stress and neuronal apoptosis through PI3K/Akt/Nrf2 pathway after intracerebral hemorrhage in mice. Oxid Med Cell Longev. 2020;2020:8864100. doi:10.1155/2020/8864100

47. Zhang XH, Weissbach H. Origin and evolution of the protein-repairing enzymes methionine sulphoxide reductases. Biol Rev Camb Philos Soc. 2008;83(3):249257. doi:10.1111/j.1469-185X.2008.00042.x

48. Wienholz F, Zhou D, Turkyilmaz Y, et al. FACT subunit Spt16 controls UVSSA recruitment to lesion-stalled RNA Pol II and stimulates TC-NER. Nucleic Acids Res. 2019;47:40114025. doi:10.1093/nar/gkz055

49. Higa M, Zhang X, Tanaka K, et al. Stabilization of Ultraviolet (UV)-stimulated scaffold protein A by interaction with ubiquitin-specific peptidase 7 is essential for transcription-coupled nucleotide excision repair. J Biol Chem. 2016;291:1377113779. doi:10.1074/jbc.M116.724658

50. Jackson E, Heidl M, Imfeld D, et al. Discovery of a highly selective MC1R agonists pentapeptide to be used as a skin pigmentation enhancer and with potential anti-aging properties. Int J Mol Sci. 2019;20:6143. doi:10.3390/ijms20246143

51. Wessler I, Kilbinger H, Bittinger F, et al. The non-neuronal cholinergic system in humans: expression, function and pathophysiology. Life Sci. 2003;72:20552061. doi:10.1016/S0024-3205(03)00083-3

52. Said ER, Nagui NA, Rashed LA, et al. Oxidative stress and the cholinergic system in non-segmental vitiligo: effect of narrow band ultraviolet b. Photodermatol Photoimmunol Photomed. 2021;37:306312. doi:10.1111/phpp.12653

53. Taieb A. Intrinsic and extrinsic pathomechanisms in vitiligo. Pigment Cell Res. 2000;13(suppl 8):4147. doi:10.1034/j.1600-0749.13.s8.9.x

54. Schallreuter KU, Wood JM, Pittelkow MR, et al. Increased monoamine oxidase A activity in the epidermis of patients with vitiligo. Arch Dermatol Res. 1996;288:1418. doi:10.1007/BF02505037

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Pigmentation and cell protection effect in melanocytes | DDDT - Dove Medical Press

OXFORD, England--(BUSINESS WIRE)--Neuro-Bio Ltd, a biotechnology company developing a first-in-class treatment for neurodegenerative disease, finds its new drug candidate effectively treats the signs of neurodegeneration in a mouse model of Alzheimers disease.

Publishing their work in Alzheimers & Dementia: Translational Research & Clinical Interventions (TRCI), Neuro-Bio researchers, in collaboration with the drug discovery company Evotec SE, UCLA, and Kings College London studied the ability of their patented drug, NBP14, to combat neurodegeneration in an established mouse model of Alzheimers disease.

Intranasal treatment for 6 weeks resulted in a marked decrease of brain amyloid and, after 14 weeks, improved cognitive performance comparable to that of normal mice. The results underscore the effectiveness of Neuro-Bios drug candidate and represent a remarkable step forward towards the treatment of Alzheimers disease in humans.

Baroness Professor Susan Greenfield, Founder and CEO of Neuro-Bio, says: By using basic neuroscientific knowledge we have identified what we believe is an underlying mechanism driving Alzheimers disease in the brain, and have developed a molecule (NBP14) to combat it. Our recent efficacy study in mouse models further validates previous work describing an erstwhile unidentified process in neurodegeneration and offers very exciting prospects for treating the disease in humans. This research should help position the drug intercepting this process, NBP14, for human clinical trials and hopefully create an entirely new era of Alzheimers therapeutics.

Professor Paul L Herrling, former Global Head of Research of Novartis Pharma and Non-Executive Director at Neuro-Bio, says: The results consistently indicate that NBP14 might interfere with the neurotoxic process that leads to neuronal degeneration in Alzheimers. This work has very exciting implications for treating Alzheimers because it is based on a strong scientific theory that hasnt yet been applied to treatment of the disease.

The UK regulator, the Medicines & Healthcare Products Regulatory Agency, has accredited NBP14 with one of their first Innovation Passports as part of a new licensing pathway that aims to reduce the time to market for innovative medicines.

NBP14 works by intercepting the process that Neuro-Bio believe could be a primary driver of neurodegeneration, the action of a brain chemical named T14 [1, 2]. In the last twenty years since it was first identified [3], evidence has become increasingly compelling that T14 plays an important role in early cell growth and normal development. However, this action can become toxic if triggered inappropriately in maturity [4] and ultimately could lead to Alzheimers disease where brain levels of T14 are shown in the current paper to reflect degree of degeneration.

Inactivation of T14 could potentially serve as a treatment for Alzheimers by halting the early advance of cell damage occurring first in primarily vulnerable cells deep the brain. Initially identified by the neurologist Martin Rossor back in 1981 [5], these primarily vulnerable cells form a kind of central hub in the brain, extending up from the top of the spinal cord. A key feature is that they are the first to display a pathology early in neurodegeneration [6].

Neuro-Bio believes that detection and measurement of T14 could be developed as a blood test or skin biopsy to identify the occurrence of the degenerative process during the window of ten to twenty years that typically occurs before symptoms start. If NBP14 proves effective in human trials, it could become a routine, home-administered nasal spray to halt neurodegeneration before any symptoms appear.

No harmful side effects at the active dose in the disease model were observed with NBP14 during the efficacy study. Neuro-Bio plan to take the drug to clinical Phase I trials as soon as possible.

Dr Gregory Cole, Professor of Medicine and Neurology at UCLA and Associate Director of the UCLA Alzheimer's Center, says: I've specialized in Alzheimers disease research and worked for real treatments in UCLAs Alzheimer program since 1994. Along with my colleagues around the world, weve all witnessed hundreds of failed drug trials based on existing theories and we are ready for truly new approaches. NBP14 has distinct advantages over other drug candidates and I am happy to work with this team at Neuro-Bio and share in their success.

ENDS

Notes to Editors

About Neuro-Bio

Neuro-Bio is a privately-owned biotech out of Oxford University with a focus on developing a first-in-class effective treatment for neurodegenerative disease. The company has discovered a novel 14 amino acid bioactive peptide (T14) derived from the C terminus of acetylcholinesterase (AChE). T14 is neurotoxic in the mature brain and published data shows it to be a potential key driver of neurodegeneration.

Based on almost 40 years of research by Professor Baroness Greenfield at Oxford University, Neuro-Bio Ltd was incorporated in 2013, when seed funding enabled the first patent filing on chemical composition of matter for neuroprotection.

For more information, visit, http://www.neuro-bio.com

About NBP14

NBP14 is a cyclated peptide, ie a structurally modified form of T14 itself, bent into a circle such that it is inactive at the target (an allosteric site on the alpha-7 receptor). By occupying this target, it will displace its naturally occurring, potentially toxic counterpart from the crucial site of action, thereby protecting against further degeneration. Hence, as reported in the paper, subsequent downstream effects will be blocked such as the production of amyloid in the hippocampus.

References

[1] Garcia-Rates S, Morrill P, Tu H, Pottiez G, Badin AS, Tormo-Garcia C, et al. (I) Pharmacological profiling of a novel modulator of the alpha7 nicotinic receptor: Blockade of a toxic acetylcholinesterase-derived peptide increased in Alzheimer brains. Neuropharmacology. 2016;105:487-99.[2] Brai E, Simon F, Cogoni A, Greenfield SA. Modulatory Effects of a Novel Cyclized Peptide in Reducing the Expression of Markers Linked to Alzheimer's Disease. Front Neurosci. 2018;12:362.[3] Greenfield S, Vaux DJ. Parkinson's disease, Alzheimer's disease and motor neurone disease: identifying a common mechanism. Neuroscience. 2002;113:485-92.[4] Day T, Greenfield SA. Bioactivity of a peptide derived from acetylcholinesterase in hippocampal organotypic cultures. Exp Brain Res. 2004;155:500-8.[5] Rossor MN. Parkinson's disease and Alzheimer's disease as disorders of the isodendritic core. Br Med J (Clin Res Ed). 1981;283:1588-90.[6] Theofilas P, Dunlop S, Heinsen H, Grinberg LT. Turning on the Light Within: Subcortical Nuclei of the Isodentritic Core and their Role in Alzheimer's Disease Pathogenesis. J Alzheimers Dis. 2015;46:17-34.

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Novel Drug From Neuro-Bio Effective in a Mouse Model of Alzheimer's Disease - Business Wire