Sophorolipid inhibits histamine‐induced itch by decreasing PLC/IP3R signaling pathway activation and modulating TRPV1 activity

Rui-Qi Xu ¹, Ling Ma ², Timson Chen ³, Wei-Xiong Zhang ³, Kuan Chang ⁴, Jing Wang ⁵

• ¹Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi, 214122, China.
• ²Adolph Innovation Laboratory, Guangzhou Degu Personal Care Products Co., Ltd., Guangzhou, 510000, China. maling@adolph.cn.
• ³Adolph Innovation Laboratory, Guangzhou Degu Personal Care Products Co., Ltd., Guangzhou, 510000, China.
• ⁴Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi, 214122, China. changkuan@jiangnan.edu.cn.
• ⁵Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi, 214122, China. jingwang@jiangnan.edu.cn.

Biosurfactants are attracting much interest due to their potential application as therapeutic agents in the medical and cosmetic field. Previous studies have demonstrated that biosurfactant such as sophorolipid (SL) exhibits immunomodulatory effects. In this article, we found the potential of sophorolipid for inhibiting histamine‐induced itch and preliminarily explored its molecular basis. First, behavioral tests indicated that SL can remit the histamine‐induced scratching behaviors of mice. Second, SL can suppress the the calcium influx induced by histamine, HTMT and VUF8430 in HaCaT cells. RT‐PCR analysis showed that the histamine‐induced upregulation of mRNA levels of phospholipase Cγ1, 1,4,5‐trisphosphate receptor (IP3R), and protein kinase Cα can be inhibted by SL, suggesting that SL may impede the PLC/IP3R signaling pathway activated by histamine. In further tests, the capsaicin‐induced calcium influx can also be inhibited by SL. The and molecular docking analysis indicated that SL acts as an inhibitor of transient receptor potential vanilloid‐1 (TRPV1) activation to decrease calcium influx against stimuli. In summary, these results revealed that SL may inhibit histamine‐induced itch by decreasing PLC/IP3R signaling pathway activation and modulating TRPV1 activity. This paper indicates that SL can be a useful treatment for histamine‐dependent itch.immunofluorescence

Activation of the Melastatin-Related Cation Channel TRPM3 by D-erythro-Sphingosine

Mol Pharmacol. 2005 Mar;67(3):798-805. doi: 10.1124/mol.104.006734. Epub 2004 Nov 18.

Christian Grimm, Robert Kraft, Gunter Schultz, and Christian Harteneck

Abstract

TRPM3, a member of the melastatin-like transient receptor potential channel subfamily (TRPM), is predominantly expressed in human kidney and brain. TRPM3 mediates spontaneous Ca2+ entry and nonselective cation currents in transiently transfected human embryonic kidney 293 cells. Using measurements with the Ca2+-sensitive fluorescent dye fura-2 and the whole-cell patch-clamp technique, we found that D-erythro-sphingosine, a metabolite arising during the de novo synthesis of cellular sphingolipids, activated TRPM3. Other transient receptor potential (TRP) channels tested [classic or canonical TRP (TRPC3, TRPC4, TRPC5), vanilloid-like TRP(TRPV4, TRPV5, TRPV6), and melastatin-like TRP (TRPM2)] did not significantly respond to application of sphingosine. Sphingosine-induced TRPM3 activation was not mediated by inhibition of protein kinase C, depletion of intracellular Ca2+ stores, and intracellular conversion of sphingosine to sphingosine-1-phosphate. Although sphingosine-1-phosphate and ceramides had no effect, two structural analogs of sphingosine, dihydro-D-erythro-sphingosine and N,N-dimethyl-D-erythro-sphingosine, also activated TRPM3. Sphingolipids, including sphingosine, are known to have inhibitory effects on a variety of ion channels. Thus, TRPM3 is the first ion channel activated by sphingolipids.

Cell Immunol 2021 Oct;368:104422. doi: 10.1016/j.cellimm.2021.104422. Epub 2021 Aug 8.

Meesum Syed ¹, Ananth K Kammala ¹, Brianna Callahan ¹, Carole A Oskeritzian ², Hariharan Subramanian ³Affiliations expand

• PMID: 34399172
• PMCID: PMC8428143
• DOI: 10.1016/j.cellimm.2021.104422

Abstract

MAS related G-protein coupled receptor X2 (MRGPRX2) is a G-protein coupled receptor (GPCR) expressed in human mast cells that has been implicated to play an important role in causing pseudo-allergic reactions as well as exacerbating inflammation during asthma and other allergic diseases. Lactic acid, a byproduct of glucose metabolism, is abundantly present in inflamed tissues and has been shown to regulate functions of several immune cells. Because the endogenous ligands for MRGPRX2 (substance P and LL-37) are elevated during pathologic conditions, such as cancer and asthma, and given that lactic acid levels are also enhanced in these patients, we explored the role of lactic acid in regulating mast cells response via MRGPRX2 and MrgprB2, the mouse orthologue of the human receptor. We found that lactic acid suppressed both the early (Ca²⁺ mobilization and degranulation) and late (chemokine/cytokine release) phases of mast cell activation; this data was confirmed in LAD2, human skin and mouse peritoneal mast cells. In LAD2 cells, the reduction in degranulation and chemokine/cytokine production mediated by lactic acid was dependent on pH. In agreement with our in vitro studies, lactic acid also reduced passive systemic anaphylaxis to compound 48/80 (a known MRGPRX2/MrgprB2 ligand) and skin inflammation in a mouse model of rosacea that is dependent on MrgprB2 expression on skin mast cells. Our data thus suggest that lactic acid may serve to inhibit mast cell-mediated inflammation during asthma and reduce immune response during cancer by affecting mast cell activation through MRGPRX2.

Keywords: Lactic acid; MAS-related G-protein coupled receptor-X2 (MRGPRX2); Mast cells; MrgprB2; Pseudo-allergic reactions.

Copyright © 2021 Elsevier Inc. All rights reserved.

MrgprA3-expressing pruriceptors drive pruritogen-induced alloknesis through mechanosensitive Piezo2 channel

Ping Lu ¹, Yonghui Zhao ², Zili Xie ², Huan Zhou ³, Xinzhong Dong ⁴, Gregory F Wu ⁵, Brian S Kim ⁶, Jing Feng ⁷, Hongzhen Hu ⁸

Abstract

Although touch and itch are coded by distinct neuronal populations, light touch also provokes itch in the presence of exogenous pruritogens, resulting in a phenomenon called alloknesis. However, the cellular and molecular mechanisms underlying the initiation of pruritogen-induced mechanical itch sensitization are poorly understood. Here, we show that intradermal injections of histamine or chloroquine (CQ) provoke alloknesis through activation of TRPV1- and MrgprA3-expressing prurioceptors, and functional ablation of these neurons reverses pruritogen-induced alloknesis. Moreover, genetic ablation of mechanosensitive Piezo2 channel function from MrgprA3-expressing prurioceptors also dampens pruritogen-induced alloknesis. Mechanistically, histamine and CQ sensitize Piezo2 channel function, at least in part, through activation of the phospholipase C (PLC) and protein kinase C-δ (PKCδ) signaling. Collectively, our data find a TRPV1⁺/MrgprA3⁺ prurioceptor-Piezo2 signaling axis in the initiation of pruritogen-induced mechanical itch sensitization in the skin.

Keywords: CP: Neuroscience; PKCδ; PLC; Piezo2; alloknesis; pruriceptor; pruritogens.

SpiCee: A Genetic Tool for Subcellular and Cell-Specific Calcium Manipulation

Oriol Ros,1,6 Sarah Baudet,1,6 Yvrick Zagar,1 Karine Loulier,1 Fiona Roche,1 Sandrine Couvet,1 Alain Aghaie,2
Melody Atkins,3 Alice Louail,1 Christine Petit,2,4 Christine Metin,3 Yves Mechulam,5 and Xavier Nicol1,7,*

Abstract

Calcium is a second messenger crucial to a myriad of cellular processes ranging from regulation of metabolism and cell survival to vesicle release and motility. Current strategies to directly manipulate endogenous calcium signals lack cellular and subcellular specificity. We introduce SpiCee, a versatile and genetically encoded chelator combining low- and high-affinity sites for calcium. This scavenger enables altering endogenous calcium signaling and functions in single cells in vitro and in vivo with biochemically controlled subcellular resolution. SpiCee paves the way to investigate local calcium signaling in vivo and directly manipulate
this second messenger for therapeutic use.

Presenter: Ju hee Ryu

Nat Commun. 2023 Apr 29;14(1):2478. doi: 10.1038/s41467-023-38209-x.

Yuanbo Jia ^# 1 2, Jiahui Hu ^# 2 3, Keli An ^1 2, Qiang Zhao ^2 3, Yang Dang ^2 3, Hao Liu ^1 2, Zhao Wei ^1 2, Songmei Geng ⁴, Feng Xu ^5 6

• PMID: 37120459
• PMCID: PMC10148840
• DOI: 10.1038/s41467-023-38209-x

Abstract

Atopic dermatitis (AD) is a chronic skin disease caused by skin immune dyshomeostasis and accompanied by severe pruritus. Although oxidative stress and mechanical scratching can aggravate AD inflammation, treatment targeting scratching is often overlooked, and the efficiency of mechano-chemically synergistic therapy remains unclear. Here, we find that enhanced phosphorylation of focal adhesion kinase (FAK) is associated with scratch-exacerbated AD. We then develop a multifunctional hydrogel dressing that integrates oxidative stress modulation with FAK inhibition to synergistically treat AD. We show that the adhesive, self-healing and antimicrobial hydrogel is suitable for the unique scratching and bacterial environment of AD skin. We demonstrate that it can scavenge intracellular reactive oxygen species and reduce mechanically induced intercellular junction deficiency and inflammation. Furthermore, in mouse AD models with controlled scratching, we find that the hydrogel alleviates AD symptoms, rebuilds the skin barrier, and inhibits inflammation. These results suggest that the hydrogel integrating reactive oxygen species scavenging and FAK inhibition could serve as a promising skin dressing for synergistic AD treatment.

Sphingosylphosphorylcholine is an activator of transglutaminase activity in human keratinocytes

Kazuhiko Higuchi,* Makoto Kawashima,* Yutaka Takagi,† Hidehiko Kondo,† Yukihiro Yada,† Yoshiaki Ichikawa,† and Genji Imokawa1,†

Abstract

Abstract We characterize functional roles of a newly discovered chemical, sphingosylphosphorylcholine (SPC), in the epidermis by elucidating the biological effect of SPC on human keratinocytes in culture. The intracellular calcium level of human keratinocytes was increased by incubation with SPC, but not with sphingosine (SS) or sphingomyelin(SM). The addition of SPC, sphingosine 1-phosphate (SSP), or SS to human keratinocytes at 10 mM concentrations also
significantly suppressed DNA synthesis, and SPC, but not SSP, or SS increased the activities of membrane-bound and
soluble transglutaminases (TGases). Reverse transcription polymerase chain reaction (RT-PCR) of TGase transcripts revealed that SPC treatment at 10 mM concentrations increased the expression of TGase 1 mRNA. The increased activity of soluble TGase was accompanied by the concomitant activation of cathepsin D as revealed by the increased
ratio of mature active form to inactive intermediate form of the protease. Pretreatment of human keratinocytes with
pepstatin, a protease inhibitor, blocked the increase in soluble TGase activity induced by treatment with SPC. Consistently, SPC treatment at 1–10 µM concentrations stimulated the cornified envelope formation. These findings
suggest that SPC plays an important role in the altered keratinization process of epidermis in skin diseases with high
expression of sphingomyelin deacylase, such as atopic dermatitis.—Higuchi, K., M. Kawashima, Y. Takagi, H. Kondo, Y.
Yada, Y. Ichikawa, and G. Imokawa. Sphingosylphosphorylcholine is an activator of transglutaminase activity in human
keratinocytes. J. Lipid Res. 2001. 42: 1562–1570.

Supplementary key words sphingomyelin deacylase • atopic dermatitis • sphingolipid • ceramide

Hair growth promotion by Necrostatin‐1s

Mei Zheng1, Nahyun Choi1, YaeJi Jang1, Da Eun Kwak2, YoungSoo Kim2, Won‐Serk Kim3,Sang Ho Oh4 & Jong‐Hyuk Sung2*

1STEMORE Co. Ltd, Incheon, South Korea. 2College of Pharmacy, Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahakro, Yeonsu-gu, Incheon 21983, South Korea. 3Department of Dermatology, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul 03181, South Korea. 4Department of Dermatology, Severance Hospital and Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul 03722, South Korea.*email: brian99@yonsei.ac.kr

Necrostatins (Necs) have been developed as a receptor‐interacting protein kinase 1 (RIPK1) inhibitor, thus inhibiting necroptosis. In this current study, we have investigated the possible involvement of necroptosis in the hair cycle regulation and further examined its underlying molecular mechanisms. Diverse RIPK1/3 inhibitors and siRNA were tested in the human outer‐root sheath (ORS) cells and animal models. The expression and hair cycle‐dependent expression of RIPK 1, respectively, were investigated in the hair follicles (HF) of human, pig, and the mouse. Resulting from the experiment, Nec‐1s was most effective in the hair growth promotion among several inhibitors. Nec‐1s induced the ORS cell proliferation and migration, and increased the HF length in mouse and pig organ cultures.

In addition, it accelerated the telogen‐to‐anagen transition and elongated the anagen period in
the mouse model. Both apoptosis and necroptosis were detected in hair cycle. RIPK1 and RIPK3 were highly expressed in ORS cells during the hair regression period. Nec‐1s upregulated the mRNA expression of Wnt3a and Wnt5b, and the activity of β‐catenin. Collectively, Nec‐1s promotes hair growth through inhibiting necroptosis and activating the Wnt/β‐catenin pathway. Necroptosis is involved in hair cycle regression, and Nec‐1s is a promising target for hair‐loss treatment.

Mechanisms and treatments of neuropathic itch in a mouse model of lymphoma

Ouyang Chen , Qianru He, Qingjian Han, Kenta Furutani, Yun Gu, Madelynne Olexa, Ru-Rong Ji

DOI: 10.1172/JCI160807

Abstract

Our understanding of neuropathic itch is limited due to a lack of relevant animal models. Patients with cutaneous T cell lymphoma (CTCL) experience severe itching. Here, we characterize a mouse model of chronic itch with remarkable lymphoma growth, immune cell accumulation, and persistent pruritus. Intradermal CTCL inoculation produced time-dependent changes in nerve innervations in lymphoma-bearing skin. In the early phase (20 days), CTCL caused hyperinnervations in the epidermis. However, chronic itch was associated with loss of epidermal nerve fibers in the late phases (40 and 60 days). CTCL was also characterized by marked nerve innervations in mouse lymphoma. Blockade of C-fibers reduced pruritus at early and late phases, whereas blockade of A-fibers only suppressed late-phase itch. Intrathecal (i.t.) gabapentin injection reduced late-phase, but not early-phase, pruritus. IL-31 was upregulated in mouse lymphoma, whereas its receptor Il31ra was persistently upregulated in Trpv1-expressing sensory neurons in mice with CTCL. Intratumoral anti-IL-31 treatment effectively suppressed CTCL-induced scratching and alloknesis (mechanical itch). Finally, i.t. administration of a TLR4 antagonist attenuated pruritus in early and late phases and in both sexes. Collectively, we have established a mouse model of neuropathic and cancer itch with relevance to human disease. Our findings also suggest distinct mechanisms underlying acute, chronic, and neuropathic itch.

Keywords: Lymphomas; Neuroscience; Skin cancer; Therapeutics.

Presenter: Diwas Rawal

Br J Pharmacol. 2023 Mar 16. doi: 10.1111/bph.16072. Online ahead of print.

Molecular mechanisms of MrgprA3-independent activation of the transient receptor potential ion channels TRPA1 and TRPV1 by chloroquine

Tabea C Fricke ¹, Sebastian Pantke ¹, Bjarne Lüttmann ¹, Frank G Echtermeyer ¹, Christine Herzog ¹, Mirjam J Eberhardt ¹, Andreas Leffler ¹

• PMID: 36928865
• DOI: 10.1111/bph.16072

Abstract

Background and purpose: Itch associates several pathologies and is a common drug-induced side effect. Chloroquine (CQ) was reported to induce itch by activating the Mas-related G protein-coupled receptor MrgprA3 and subsequently TRPA1. In this study we demonstrate that CQ employs at least two MrgprA3-independent mechanisms to activate or sensitize TRPA1 and TRPV1.

Experimental approach: Patch Clamp and calcium-imaging were utilized to examine effects of CQ on TRPA1 and TRPV1 expressed in HEK-293T cells.

Key results: In calcium-imaging, CQ induces a concentration-dependent but MrgprA3-independent activation of TRPA1 and TRPV1. While CQ itself inhibits TRPA1 and TRPV1 in patch clamp recordings, co-application of CQ and UVA-light evokes membrane currents through both channels. This effect is inhibited by the reducing agent dithiothreitol (DTT) and reduced on mutants lacking cysteine residues accounting for reactive oxygen species (ROS)-sensitivity. The combination of CQ and UVA-light triggers an accumulation of intracellular ROS, removes fast inactivation of voltage-gated sodium currents and activates TRPV2. On the other hand, CQ is a weak base and induces intracellular alkalosis. Intracellular alkalosis can activate TRPA1 and TRPV1, and CQ applied at alkaline pH-values indeed activates both channels.

Conclusion and implications: Our data reveal novel pharmacological properties of CQ allowing activation of TRPA1 and TRPV1 via photosensitization as well as intracellular alkalosis. These findings add complexity to the commonly accepted dogma that CQ-induced itch is specifically mediated by MrgprA3 coupling to TRPA1.

Keywords: chloroquine; histamine; itch; oxidative stress; sensory neuron.