Publications

@article{InKim2023,

title = {FSLLRY-NH2, a protease-activated receptor 2 (PAR2) antagonist, activates mas-related G protein-coupled receptor C11 (MrgprC11) to induce scratching behaviors in mice},

author = {Hye In Kim and Gi Baek Lee and Da Eun Song and Babina Sanjel and Wook-Joo Lee and Won-Sik Shim},

doi = {10.1016/j.lfs.2023.121786},

issn = {0024-3205},

year  = {2023},

date = {2023-07-00},

urldate = {2023-07-00},

journal = {Life Sciences},

volume = {325},

publisher = {Elsevier BV},

abstract = {Aims: Protease-activated receptor 2 (PAR2), a type of G protein-coupled receptor (GPCR), plays a significant role in pathophysiological conditions such as inflammation. A synthetic peptide SLIGRL-NH2 (SLIGRL) can activate PAR2, while FSLLRY-NH2 (FSLLRY) is an antagonist. A previous study showed that SLIGRL activates both PAR2 and mas-related G protein-coupled receptor C11 (MrgprC11), a different type of GPCR expressed in sensory neurons. However, the impact of FSLLRY on MrgprC11 and its human ortholog MRGPRX1 was not verified. Hence, the present study aims to verify the effect of FSLLRY on MrgprC11 and MRGPRX1.



Methods: The calcium imaging technique was applied to determine the effect of FSLLRY in HEK293T cells expressing MrgprC11/MRGPRX1 or dorsal root ganglia (DRG) neurons. Scratching behavior was also investigated in wild-type and PAR2 knockout mice after injecting FSLLRY.



Key findings: It was surprisingly discovered that FSLLRY specifically activates MrgprC11 in a dose-dependent manner, but not other MRGPR subtypes. Furthermore, FSLLRY also moderately activated MRGPRX1. FSLLRY stimulates downstream pathways including Gαq/11, phospholipase C, IP3 receptor, and TRPC ion channels to evoke an increase in the intracellular calcium levels. The molecular docking analysis predicted that FSLLRY interacts with the orthosteric binding pocket of MrgprC11 and MRGPRX1. Finally, FSLLRY activated primary cultures of mouse sensory neurons, and induced scratching behaviors in mice.



Significance: The present study has revealed that FSLLRY is capable of triggering itch sensation through activation of MrgprC11. This finding highlights the importance of considering the unexpected activation of MRGPRs in future therapeutic approaches aimed at the inhibition of PAR2.},

keywords = {Calcium imaging, FSLLRY, Itch, MrgprC11, MRGPRX1, PAR2, Scratching behavior},

pubstate = {published},

tppubtype = {article}

}

@article{Lee2023,

title = {Rg3-enriched Korean red ginseng alleviates chloroquine-induced itch and dry skin pruritus in an MrgprA3-dependent manner in mice},

author = {Wook-Joo Lee and Won-Sik Shim},

doi = {10.1016/j.imr.2022.100916},

issn = {2213-4220},

year  = {2023},

date = {2023-03-00},

urldate = {2023-03-00},

journal = {Integrative Medicine Research},

volume = {12},

number = {1},

publisher = {Elsevier BV},

abstract = {Background: Previous studies have found that Korean red ginseng extract (KRG) has antipruritic effects, which can be attributed to the presence of Rg3, one of the most potent ginsenosides. Therefore, Rg3-enriched KRG extract (Rg3EKRG) is anticipated to have enhanced antipruritic effects. The present study was conducted to examine the effects of Rg3EKRG in acute chloroquine (CQ)-induced and chronic dry skin pruritus.



Methods: Calcium imaging technique was used in HE293T cells expressing MrgprA3 and TRPA1 ("MrgprA3/TRPA1") and in primary cultures of mouse dorsal root ganglia (DRG) neurons. Mouse scratching behavior tests were performed on dry skin models. To verify the altered expression of itch-related genes, real-time RNA sequencing analysis and PCR were performed on DRG sections obtained from dry skin models.



Results: Rg3EKRG suppressed CQ-induced intracellular calcium changes to a greater degree than KRG. Rg3EKRG dose-dependently inhibited CQ-induced responses in MrgprA3/TRPA1 cells. Rg3EKRG likely targeted MrgprA3 rather than TRPA1 to exert its inhibitory effect. Further, Rg3EKRG strongly inhibited the scratching behavior in mice induced by acute CQ injection. Importantly, DRG neurons obtained from dry skin mice models showed increased mRNA levels of MrgprA3, and treatment with Rg3EKRG alleviated chronic dry skin conditions and suppressed spontaneous scratching behaviors.



Conclusion: The results of the present study imply that Rg3EKRG has a stronger antipruritic effect than KRG, inhibiting both acute CQ-induced and chronic dry skin pruritus in an MrgprA3-dependent manner. Therefore, Rg3EKRG is a potential antipruritic agent that can suppress acute and chronic itching at the peripheral sensory neuronal level.},

keywords = {Dry skin, Itch, Korean Red Ginseng, MrgprA3, RNA-seq, Scratching behavior, TRPA1},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2023,

title = {Pathogenesis and Treatment of Pruritus Associated with Chronic Kidney Disease and Cholestasis},

author = {Jin-Cheol Kim and Won-Sik Shim and In-Suk Kwak and Dong-Hun Lee and Jin-Seo Park and So-Yeon Lee and Seok-Young Kang and Bo-Young Chung and Chun-Wook Park and Hye-One Kim},

doi = {10.3390/ijms24021559},

issn = {1422-0067},

year  = {2023},

date = {2023-01-00},

urldate = {2023-01-00},

journal = {IJMS},

volume = {24},

number = {2},

publisher = {MDPI AG},

abstract = {Itching is an unpleasant sensation that provokes the desire to scratch. In general, itching is caused by dermatologic diseases, but it can also be caused by systemic diseases. Since itching hampers patients’ quality of life, it is important to understand the appropriate treatment and pathophysiology of pruritus caused by systemic diseases to improve the quality of life. Mechanisms are being studied through animal or human studies, and various treatments are being tested through clinical trials. We report current trends of two major systemic diseases: chronic kidney disease and cholestatic liver disease. This review summarizes the causes and pathophysiology of systemic diseases with pruritus and appropriate treatments. This article will contribute to patients’ quality of life. Further research will help understand the mechanisms and develop new strategies in the future.},

keywords = {Cholestasis, Chronic kidney disease, Itch},

pubstate = {published},

tppubtype = {article}

}

@article{Afzal2022,

title = {Activation of serotonin receptor 2 by glucosylsphingosine can be enhanced by TRPA1 but not TRPV1: Implication of a novel glucosylsphingosine-mediated itch pathway},

author = {Ramsha Afzal and Won-Sik Shim},

doi = {10.1016/j.bbamem.2022.184014},

issn = {0005-2736},

year  = {2022},

date = {2022-11-00},

urldate = {2022-11-00},

journal = {Biochimica et Biophysica Acta (BBA) - Biomembranes},

volume = {1864},

number = {11},

publisher = {Elsevier BV},

abstract = {Glucosylsphingosine (GS) is an endogenous sphingolipid that specifically accumulates in the skin of patients with atopic dermatitis (AD). Notably, it was recently found that GS can induce itch sensation by activating serotonin receptor 2A and TRPV4 ion channels. However, it is still uncertain whether other molecules are involved in GS-induced itch sensation. Therefore, by using the calcium imaging technique, we investigated whether serotonin receptor 2 - specifically 2A and 2B - can interact with TRPV1 and TRPA1, because these are representative ion channels in the transmission of itch. As a result, it was found that GS did not activate TRPV1 or TRPA1 per se. Moreover, cells expressing both serotonin receptor 2 and TRPV1 did not show any changes in calcium responses. However, enhanced calcium responses were observed in cells expressing serotonin receptor 2 and TRPA1, suggesting a possible interaction between these two molecules. Similar synergistic effects were also observed in cells expressing serotonin receptor 2 and TRPA1, but not TRPV1. Furthermore, a phospholipase C inhibitor (U73122) and a store-operated calcium entry blocker (SKF96365) significantly reduced GS-induced responses in cells expressing both serotonin receptor 2 and TRPA1, but not with pre-treatment with a Gβγ-complex blocker (gallein). Therefore, we propose a putative novel pathway for GS-induced itch sensation, such that serotonin receptor 2 could be coupled to TRPA1 but not TRPV1 in sensory neurons.},

keywords = {5-HT2R, Calcium imaging, Glucosylsphingosine, TRPA1, TRPV1},

pubstate = {published},

tppubtype = {article}

}

@article{Adhikari2022,

title = {Caffeic acid phenethyl ester inhibits pseudo-allergic reactions via inhibition of MRGPRX2/MrgprB2-dependent mast cell degranulation},

author = {Nisha Adhikari and Won-Sik Shim},

doi = {10.1007/s12272-022-01405-2},

issn = {1976-3786},

year  = {2022},

date = {2022-09-00},

urldate = {2022-09-00},

journal = {Arch. Pharm. Res.},

volume = {45},

number = {9},

pages = {644--657},

publisher = {Springer Science and Business Media LLC},

abstract = {Mast cells play essential role in allergic reactions through the process called mast cell degranulation. Recent studies have found that a basic secretagogue compound 48/80 (C48/80) induces non-IgE-mediated mast cell degranulation via activation of human Mas-related G protein-coupled receptor X2 (MRGPRX2) and mouse MrgprB2. Although previous studies have revealed that caffeic acid (CA) and its derivatives possess anti-allergic effects via IgE-dependent manner, it is largely elusive whether these compounds have impact on MRGPRX2/MrgprB2 to exert inhibitory effects. Therefore, the present study investigated whether CA as well as its derivatives - rosmarinic acid (RA) and caffeic acid phenethyl ester (CAPE) - has the ability to inhibit the activity of MRGPRX2/MrgprB2 to evoke pseudo-allergic effects. As a result, it was found that CAPE inhibits C48/80-induced activation of MRGPRX2/MrgprB2, but neither CA nor RA showed discernible inhibition. Furthermore, the β-hexosaminidase release assay showed that CAPE inhibits mouse peritoneal mast cell degranulation in both IgE-dependent and MrgprB2-dependent manners. Additionally, mouse paw edema induced by C48/80 was dramatically suppressed by co-treatment of CAPE, suggesting that CAPE possesses a protective effect on C48/80-evoked pseudo-allergic reactions. The pretreatment of CAPE also significantly decreased scratching bouts of mice evoked by C48/80, demonstrating that CAPE also has an anti-pruritic effect. Therefore, these data implicate that CAPE can suppress pseudo-allergic reactions evoked by C48/80 via MrgprB2-dependent manner. Finally, molecular docking analysis showed that CAPE is predicted to bind to human MRGPRX2 in the region where C48/80 also binds, implying that CAPE can be a competitive inhibitor of MRGPRX2. In conclusion, it is found that CAPE has the ability to inhibit MRGPRX2/MrgprB2, leading to the prevention of mast cell degranulation and further to the alleviation of mast cell reactions. These results indicate that CAPE as a CA derivative could be developed as a new protective agent that exerts dual inhibition of mast cell degranulation mediated by IgE and MRGPRX2/MrgprB2.},

keywords = {Allergy, Calcium imaging, Mast cell, MrgprB2, MRGPRX2},

pubstate = {published},

tppubtype = {article}

}

@article{Sanjel2022,

title = {The contribution of mouse models to understanding atopic dermatitis},

author = {Babina Sanjel and Won-Sik Shim},

doi = {10.1016/j.bcp.2022.115177},

issn = {0006-2952},

year  = {2022},

date = {2022-09-00},

urldate = {2022-09-00},

journal = {Biochemical Pharmacology},

volume = {203},

publisher = {Elsevier BV},

abstract = {Atopic dermatitis (AD) is a dermatological disease accompanied by dry and cracked skin with severe pruritus. Although various therapeutic strategies have been introduced to alleviate AD, it remains challenging to cure the disorder. To achieve such a goal, understanding the pathophysiological mechanisms of AD is a prerequisite, requiring mouse models that properly reflect the AD phenotypes. Currently, numerous AD mouse models have been established, but each model has its own advantages and weaknesses. In this review, we categorized and summarized mouse models of AD and described their characteristics from a researcher’s perspective.},

keywords = {Animal model, Atopic dermatitis, Itch},

pubstate = {published},

tppubtype = {article}

}

@article{Sanjel2022b,

title = {Glucosylsphingosine evokes pruritus via activation of 5‐HT_{2A} receptor and TRPV4 in sensory neurons},

author = {Babina Sanjel and Bo‐Hyun Kim and Myung‐Hyun Song and Earl Carstens and Won‐Sik Shim},

doi = {10.1111/bph.15733},

issn = {1476-5381},

year  = {2022},

date = {2022-05-00},

urldate = {2022-05-00},

journal = {British J Pharmacology},

volume = {179},

number = {10},

pages = {2193--2207},

publisher = {Wiley},

abstract = {Background and purpose



Glucosylsphingosine (GS), an endogenous sphingolipid, is highly accumulated in the epidermis of patients with atopic dermatitis (AD) due to abnormal ceramide metabolism. More importantly, GS can evoke scratching behaviours. However, the precise molecular mechanism by which GS induces pruritus has been elusive. Thus, the present study aimed to elucidate the molecular signalling pathway of GS, especially at the peripheral sensory neuronal levels.



Experimental approach



Calcium imaging was used to investigate the responses of HEK293T cells or mouse dorsal root ganglion (DRG) neurons to application of GS. Scratching behaviour tests were also performed with wild-type and Trpv4 knockout mice.



Key results



GS activated DRG neurons in a manner involving both the 5-HT2A receptor and TRPV4. Furthermore, GS-induced responses were significantly suppressed by various inhibitors, including ketanserin (5-HT2A receptor antagonist), YM254890 (Gαq/11 inhibitor), gallein (Gβγ complex inhibitor), U73122 (phospholipase C inhibitor), bisindolylmaleimide I (PKC inhibitor) and HC067047 (TRPV4 antagonist). Moreover, DRG neurons from Trpv4 knockout mice exhibited significantly reduced responses to GS. Additionally, GS-evoked scratching behaviours were greatly decreased by pretreatment with inhibitors of either 5-HT2A receptor or TRPV4. As expected, GS-evoked scratching behaviour was also significantly decreased in Trpv4 knockout mice.



Conclusion and implications



Overall, the present study provides evidence for a novel molecular signalling pathway for GS-evoked pruritus, which utilizes both 5-HT2A receptor and TRPV4 in mouse sensory neurons. Considering the high accumulation of GS in the epidermis of patients with AD, GS could be another pruritogen in patients with AD.},

keywords = {5-HT2R, Atopic dermatitis, Calcium imaging, Dorsal root ganglia, Glucosylsphingosine, TRPV4},

pubstate = {published},

tppubtype = {article}

}

@article{Song2022,

title = {Lithocholic Acid Activates Mas-Related G Protein-Coupled Receptors, Contributing to Itch in Mice},

author = {Myung-Hyun Song and Won-Sik Shim},

doi = {10.4062/biomolther.2021.059},

issn = {2005-4483},

year  = {2022},

date = {2022-01-01},

urldate = {2022-01-01},

journal = {Biomol Ther (Seoul)},

volume = {30},

number = {1},

pages = {38--47},

publisher = {The Korean Society of Applied Pharmacology},

abstract = {The present study focused on lithocholic acid (LCA), a secondary bile acid that contributes to cholestatic pruritus. Although recent studies have found that LCA acts on MAS-related G protein-coupled receptor family member X4 (MRGPRX4) in humans, it is unclear which subtypes of MRGPRs are activated by LCA in mice since there is no precise ortholog of human MRGPRX4 in the mouse genome. Using calcium imaging, we found that LCA could activate mouse Mrgpra1 when transiently expressed in HEK293T cells. Moreover, LCA similarly activates mouse Mrgprb2. Importantly, LCA-induced responses showed dose-dependent effects through Mrgpra1 and Mrgprb2. Moreover, treatment with QWF (an antagonist of Mrgpra1 and Mrgprb2), YM254890 (Gαq inhibitor), and U73122 (an inhibitor of phospholipase C) significantly suppressed the LCA-induced responses, implying that the LCA-induced responses are indeed mediated by Mrgpra1 and Mrgprb2. Furthermore, LCA activated primary cultures of mouse sensory neurons and peritoneal mast cells, suggesting that Mrgpra1 and Mrgprb2 contribute to LCA-induced pruritus. However, acute injection of LCA did not induce noticeable differences in scratching behavior, implying that the pruritogenic role of LCA may be marginal in non-cholestatic conditions. In summary, the present study identified for the first time that LCA can activate Mrgpra1 and Mrgprb2. The current findings provide further insight into the similarities and differences between human and mouse MRGPR families, paving a way to understand the complex roles of these pruriceptors.},

keywords = {Bile acid, Calcium imaging, Itch, Lithocholic acid, MrgprA1, MrgprB2, MRGPRX4},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2022,

title = {Anoctamin 1/TMEM16A in pruritoceptors is essential for Mas-related G protein receptor–dependent itch},

author = {Hyesu Kim and Hyungsup Kim and Hawon Cho and Byeongjun Lee and Huan-Jun Lu and Kyungmin Kim and Sooyoung Chung and Won-Sik Shim and Young Kee Shin and Xinzhong Dong and John N. Wood and Uhtaek Oh},

doi = {10.1097/j.pain.0000000000002611},

issn = {1872-6623},

year  = {2022},

date = {2022-00-00},

urldate = {2022-00-00},

journal = {PAIN},

volume = {163},

number = {11},

pages = {2172--2184},

publisher = {Ovid Technologies (Wolters Kluwer Health)},

abstract = {Abstract

          Itch is an unpleasant sensation that evokes a desire to scratch. Pathologic conditions such as allergy or atopic dermatitis produce severe itching sensation. Mas-related G protein receptors (Mrgprs) are receptors for many endogenous pruritogens. However, signaling pathways downstream to these receptors in dorsal root ganglion (DRG) neurons are not yet understood. We found that anoctamin 1 (ANO1), a Ca2+-activated chloride channel, is a transduction channel mediating Mrgpr-dependent itch signals. Genetic ablation of Ano1 in DRG neurons displayed a significant reduction in scratching behaviors in response to acute and chronic Mrgpr-dependent itch models and the epidermal hyperplasia induced by dry skin. In vivo Ca2+ imaging and electrophysiological recording revealed that chloroquine and other agonists of Mrgprs excited DRG neurons via ANO1. More importantly, the overexpression of Ano1 in DRG neurons of Ano1-deficient mice rescued the impaired itching observed in Ano1-deficient mice. These results demonstrate that ANO1 mediates the Mrgpr-dependent itch signaling in pruriceptors and provides clues to treating pathologic itch syndromes.},

keywords = {Anoctamin 1, Itch, MrgprA3, MrgprC11},

pubstate = {published},

tppubtype = {article}

}

@article{Lee2021,

title = {Cutaneous Neuroimmune Interactions of TSLP and TRPV4 Play Pivotal Roles in Dry Skin-Induced Pruritus},

author = {Wook-Joo Lee and Won-Sik Shim},

doi = {10.3389/fimmu.2021.772941},

issn = {1664-3224},

year  = {2021},

date = {2021-12-02},

urldate = {2021-12-02},

journal = {Front. Immunol.},

volume = {12},

publisher = {Frontiers Media SA},

abstract = {Dry skin is a symptom of skin barrier dysfunction that evokes pruritus; however, the cutaneous neuroimmune interactions underlying dry skin-induced pruritus remain unclear. Therefore, we aimed to elucidate the mechanisms underlying dry skin-induced pruritus. To this end, an acetone/ethanol/water (AEW)-induced mouse model of dry skin was used in this study. We observed that the production of thymic stromal lymphopoietin (TSLP) significantly increased in the keratinocytes of AEW mice. Importantly, treatment with an antagonist of transient receptor potential cation channel subfamily V member 4 (TRPV4), HC067047, ameliorated dry skin conditions in AEW mice. The symptoms of dry skin were significantly reduced in Trpv4 knockout (KO) mice following treatment with AEW. The increase in the intracellular calcium levels by TSLP in the dorsal root ganglia (DRG) of Trpv4 KO mice was also significantly attenuated. The spontaneous scratching bouts were significantly decreased in both the HC067047-treated and Trpv4 KO AEW mice. Importantly, the TSLP-dependent release of tryptase from the mast cells was significantly reduced in both the HC067047-treated mice and Trpv4 KO AEW mice. Notably, inhibition of the TSLP-induced signaling pathway in DRG selectively reduced the spontaneous scratching bouts in AEW mice. Overall, the results demonstrated that the cutaneous neuroimmune interactions of TSLP and TRPV4 play pivotal roles in dry skin-induced pruritus.},

keywords = {Calcium imaging, Dorsal root ganglia, Dry skin, Itch, TSLP},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2021,

title = {Activation of Transient Receptor Potential Vanilloid-3 Channels in Keratinocytes Induces Pruritus in Humans},

author = {J Kim and H Kim and W Shim and I Kwak and B Chung and S Kang and C Park and H Kim},

doi = {10.2340/00015555-3855},

issn = {1651-2057},

year  = {2021},

date = {2021-08-00},

urldate = {2021-08-00},

journal = {Acta Derm Venereol},

volume = {101},

number = {8},

publisher = {Medical Journals Sweden AB},

abstract = {Carvacrol, a natural transient receptor potential vanilloid-3 activator, has been reported to cause pruritus in mice. This study aimed to evaluate the effects of carvacrol and various antipruritic agents in humans. A stimulation test with carvacrol, β-alanine, and histamine was performed. After application of the pruritic solutions, the skin was stimulated with pinpricks. In inhibition test A, Forsythia suspensa extract, containing forsythoside B (a transient receptor potential vanilloid-3 inhibitor), was applied by pricking prior to stimulation with pruritogens. In inhibition test B, olopatadine solution, tacrolimus ointment, and Scutellaria baicalensis root extract were applied, and carvacrol was applied to the same region. Carvacrol induces moderate pruritus in humans. The pruritus was relieved by Forsythia suspensa extract and olopatadine solution after 20 min of application and by tacrolimus ointment and Scutellaria baicalenis extract after 24 h of application. These results suggest that carvacrol is a pruritogen in humans, and that carvacrol-induced pruritus is inhibited by various antipruritic agents.},

keywords = {Itch, TRPV3, TSLP},

pubstate = {published},

tppubtype = {article}

}

@article{Sanjel2020,

title = {Recent advances in understanding the molecular mechanisms of cholestatic pruritus: A review},

author = {Babina Sanjel and Won-Sik Shim},

doi = {10.1016/j.bbadis.2020.165958},

issn = {0925-4439},

year  = {2020},

date = {2020-12-00},

urldate = {2020-12-00},

journal = {Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease},

volume = {1866},

number = {12},

publisher = {Elsevier BV},

abstract = {Cholestasis, a condition characterized by an abnormal decrease in bile flow, is accompanied by various symptoms such as pruritus. Although cholestatic pruritus is a prominent condition, its precise mechanisms have largely been elusive. Recently, advancements have been made for understanding the etiology and pathogenesis of cholestatic pruritus. The current review therefore focuses on summarizing the overall progress made in the elucidation of its molecular mechanisms. We have reviewed the available animal models on cholestasis to compare the differences between them, characterized potential pruritogens involved in cholestatic pruritus, and have summarized the receptor and ion channels implicated in the condition. Finally, we have discussed the available treatment options for alleviation of cholestatic pruritus. As our understanding of the mechanisms of cholestatic pruritus deepens, novel strategies to cure this condition are awaited.},

keywords = {Animal model, Cholestasis, Itch},

pubstate = {published},

tppubtype = {article}

}

@article{Yum2020,

title = {Development of a Novel Blue Fluorescent Gene-encoded Calcium Indicator Modified from GCaMP3},

author = {Chorok Yum and Taekyoung Ahn and Won-Sik Shim},

doi = {10.1007/s10895-020-02606-y},

issn = {1573-4994},

year  = {2020},

date = {2020-12-00},

urldate = {2020-12-00},

journal = {J Fluoresc},

volume = {30},

number = {6},

pages = {1287--1293},

publisher = {Springer Science and Business Media LLC},

abstract = {Intracellular calcium can be monitored by various calcium-specific fluorescent dyes including gene-encoded calcium indicators (GECI). GCaMP is a widely-used GECI that emits green fluorescence proportional to the level of intracellular calcium. However, since many tagging proteins also emit green fluorescence, GCaMP cannot be used with another green fluorescent protein. Therefore, it would be ideal to develop a GECI that has a distinct color profile other than green. In this regard, we developed a novel blue fluorescentcalcium indicator modified from GCaMP called Ser222-Ala229-Cys330-BCaMP3. Specifically, a simple threonine to histidine substitution to a green fluorescent Cys330-GCaMP3 successfully changed its fluorescence to blue (Cys330-BCaMP3, B for blue). Furthermore, a couple of additional amino acid substitutions resulted in more enhanced blue fluorescence intensity. Among other Cys330-BCaMP3 variants, it was found that Ser222-Ala229-Cys330-BCaMP3 exhibited the strongest blue fluorescence intensity. When Ser222-Ala229-Cys330-BCaMP3 was co-expressed with TRPA1 – a non-selective cation channel – in HEK293T cells, it showed moderate bluefluorescence. One of the drawbacks of Ser222-Ala229-Cys330-BCaMP3 was that the fluorescence intensity was not enough when cellswere cultured under 37°C. However, this limitation was circumvented by lowering cell culture temperature to 28°C, allowing muchmore enhanced blue fluorescence. Although Ser222-Ala229-Cys330-BCaMP3 mandates further optimization, the present study hasfound a promising blue fluorescent GECI that is derived from GCaMP3.},

keywords = {Calcium imaging, GCaMP3, GFP},

pubstate = {published},

tppubtype = {article}

}

@article{Choi2020,

title = {Crotamiton, an Anti-Scabies Agent, Suppresses Histamine- and Chloroquine-Induced Itch Pathways in Sensory Neurons and Alleviates Scratching in Mice},

author = {Da-Som Choi and Yeounjung Ji and Yongwoo Jang and Wook-Joo Lee and Won-Sik Shim},

doi = {10.4062/biomolther.2020.063},

issn = {2005-4483},

year  = {2020},

date = {2020-11-01},

urldate = {2020-11-01},

journal = {Biomolecules & Therapeutics},

volume = {28},

number = {6},

pages = {569--575},

publisher = {The Korean Society of Applied Pharmacology},

abstract = {Crotamiton is an anti-scabies drug, but it was recently found that crotamiton also suppresses non-scabietic itching in mice. However, the underlying mechanism is largely unclear. Therefore, aim of the study is to investigate mechanisms of the anti-pruritic effect of crotamiton for non-scabietic itching. Histamine and chloroquine are used as non-scabietic pruritogens. The effect of crotamiton was identified using fluorometric intracellular calcium assays in HEK293T cells and primary cultured dorsal root ganglion (DRG) neurons. Further in vivo effect was evaluated by scratching behavior tests. Crotamiton strongly inhibited histamine-induced calcium influx in HEK293T cells, expressing both histamine receptor 1 (H1R) and transient receptor potential vanilloid 1 (TRPV1), as a model of histamine-induced itching. Similarly, it also blocked chloroquine-induced calcium influx in HEK293T cells, expressing both Mas-related G-protein-coupled receptor A3 (MRGPRA3) and transient receptor potential A1 (TRPA1), as a model of histamine-independent itching. Furthermore, crotamiton also suppressed both histamine- and chloroquine-induced calcium influx in primary cultures of mouse DRG. Additionally, crotamiton strongly suppressed histamine- and chloroquine-induced scratching in mice. Overall, it was found that crotamiton has an anti-pruritic effect against non-scabietic itching by histamine and chloroquine. Therefore, crotamiton may be used as a general anti-pruritic agent, irrespective of the presence of scabies.},

keywords = {Calcium imaging, Chloroquine, H1R, Histamine, Itch, MrgprA3, TRPA1, TRPV1},

pubstate = {published},

tppubtype = {article}

}

@article{Sanjel2019,

title = {BAM8-22 and its receptor MRGPRX1 may attribute to cholestatic pruritus},

author = {Babina Sanjel and Han-Joo Maeng and Won-Sik Shim},

doi = {10.1038/s41598-019-47267-5},

issn = {2045-2322},

year  = {2019},

date = {2019-12-00},

urldate = {2019-12-00},

journal = {Sci Rep},

volume = {9},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {Pruritus is an unexpected symptom observed in cholestasis and its mechanism is still unclear. Here, we show that bovine adrenal medulla (BAM) 8–22, an endogenous itch-inducing peptide, could be involved in cholestatic pruritus. It was found that bile duct ligation (BDL) mice, an obstructive cholestasis model, showed increased spontaneous scratching behaviour. Importantly, the mRNA level of proenkephalin, a precursor polypeptide of BAM8-22, was significantly increased in the skin of BDL mice. Furthermore, the mRNA level of Mrgprx1, which encodes a receptor for BAM8-22, was significantly increased in the dorsal root ganglia (DRG) of BDL mice. This was further confirmed by elevation of intracellular calcium levels upon BAM8-22 treatment in primarily-cultured DRG neurons. In addition, BDL mice showed augmented scratching behaviour by BAM8-22, indicating enhanced activity of MRGPRX1. Moreover, the skin homogenate of BDL mice induced elevation of intracellular calcium levels through MRGPRX1. Finally, among the various bile acids, chenodeoxycholic acid significantly increased proenkephalin transcription in a human keratinocyte cell line (HaCaT). In conclusion, cholestatic pruritus could be attributed in part to enhanced action of both BAM8-22 in the skin and its receptor MRGPRX1 in sensory neurons.},

keywords = {Animal model, Calcium imaging, Cholestasis, Dorsal root ganglia, MrgprC11, MRGPRX1, Scratching behavior},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2019,

title = {Extracts of the leaves of Pyrus ussuriensis Maxim. Alleviate itch sensation via TSLP-dependent manner in mouse models of atopic dermatitis},

author = {Bo Hyun Kim and Wook-Joo Lee and Babina Sanjel and Kyohee Cho and Youn Kyoung Son and Hye Yoon Park and Sun Yeou Kim and Won-Sik Shim},

doi = {10.1016/j.physbeh.2019.112624},

issn = {0031-9384},

year  = {2019},

date = {2019-10-00},

urldate = {2019-10-00},

journal = {Physiology & Behavior},

volume = {210},

publisher = {Elsevier BV},

abstract = {Pyrus ussuriensis Maxim. commonly known as “Sandolbae” in Korean is a pear tree widely distributed across East Asia. Recent studies indicate that P. ussuriensis Maxim. leaves (PUL) have antipruritic effects. This study aimed to determine the effects of PUL extract and its fractions in decreasing the itch sensation and skin lesions in two distinct animal models of atopic dermatitis (AD) induced by dinitrofluorobenzene (DNFB) or house dust mite(HDM). Our results showed that the total ethanol extract of PUL decreased the scratching behavior in mice withDNFB- and HDM-induced AD. Moreover, the ethyl acetate fraction of PUL significantly improved the overallcondition of the mice with AD induced by HDM. Further, we used HEK293T cells that express receptors and ion channels for thymic stromal lymphopoietin (TSLP), a potent pruritogen for AD, to determine the mechanismsunderlying the antipruritic effects of PUL extract/fractions. Specific subfractions of the PUL strongly inhibited the increase in calcium levels induced by TSLP. In addition, the specific subfraction of PUL inhibited the TSLP-induced increase in calcium levels in cultured mouse dorsal root ganglia neurons. Thus, our results showed thatthe PUL extract could be effective for alleviating pruritus, and the antipruritic effects were exerted probably viathe inhibition of the TSLP pathway in peripheral sensory neurons governing the itch sensation in AD.},

keywords = {Animal model, Atopic dermatitis, Calcium imaging, Itch, Scratching behavior, TSLP},

pubstate = {published},

tppubtype = {article}

}

@article{Lee2018,

title = {Korean Red Ginseng extract and ginsenoside Rg3 have anti-pruritic effects on chloroquine-induced itch by inhibition of MrgprA3/TRPA1-mediated pathway},

author = {Wook-Joo Lee and Young-Sik Kim and Won-Sik Shim},

doi = {10.1016/j.jgr.2017.05.004},

issn = {1226-8453},

year  = {2018},

date = {2018-10-00},

urldate = {2018-10-00},

journal = {Journal of Ginseng Research},

volume = {42},

number = {4},

pages = {470--475},

publisher = {Elsevier BV},

abstract = {Background: It was previously found that Korean Red Ginseng water extract (KRGE) inhibits the histamine-induced itch signaling pathway in peripheral sensory neurons. Thus, in the present study, we investigated whether KRGE inhibited another distinctive itch pathway induced by chloroquine (CQ); a representative histamine-independent pathway mediated by MrgprA3 and TRPA1. Methods: Intracellular calcium changes were measured by the calcium imaging technique in theHEK293T cells transfected with both MrgprA3 and TRPA1 (“MrgprA3/TRPA1”), and in primary culture ofmouse dorsal root ganglia (DRGs). Mouse scratching behavior tests were performed to verify proposed antipruritic effects of KRGE and ginsenoside Rg3.Results: CQ-induced Ca2þ influx was strongly inhibited by KRGE (10 mg/mL) in MrgprA3/TRPA1, and notablyginsenoside Rg3 dose-dependently suppressed CQ-induced Ca2þ influx in MrgprA3/TRPA1. Moreover, bothKRGE (10 mg/mL) and Rg3 (100 mM) suppressed CQ-induced Ca2þ influx in primary culture of mouse DRGs,indicating that the inhibitory effect of KRGE was functional in peripheral sensory neurons. In vivo tests revealed that not only KRGE (100 mg) suppressed CQ-induced scratching in mice [bouts of scratching:274.0  51.47 (control) vs. 104.7  17.39 (KRGE)], but also Rg3 (1.5 mg) oral administration significantly reduced CQ-induced scratching as well [bouts of scratching: 216.8  33.73 (control) vs.115.7  20.94 (Rg3)]. Conclusion: The present study verified that KRGE and Rg3 have a strong antipruritic effect against CQ-induced itch. Thus, KRGE is as a promising antipruritic agent that blocks both histamine-dependent and -independent itch at peripheral sensory neuronal levels.},

keywords = {Calcium imaging, Chloroquine, Korean Red Ginseng, MrgprA3, Scratching behavior, TRPA1},

pubstate = {published},

tppubtype = {article}

}

@article{Ji2018,

title = {Different perception levels of histamine-induced itch sensation in young adult mice},

author = {Yeounjung Ji and Yongwoo Jang and Wook Joo Lee and Young Duk Yang and Won-Sik Shim},

doi = {10.1016/j.physbeh.2018.02.015},

issn = {0031-9384},

year  = {2018},

date = {2018-05-00},

urldate = {2018-05-00},

journal = {Physiology & Behavior},

volume = {188},

pages = {188--193},

publisher = {Elsevier BV},

abstract = {Itch is an unpleasant sensation that evokes behavioral responses such as scratching the skin. Interestingly, it isconceived that the perception of itch sensation is influenced by age. Indeed, accumulating evidence supports theidea that even children or younger adults show distinctive itch sensation depending on age. This evidence implies the presence of a mechanism that regulates the perception of itch sensation in an age-dependent fashion. Therefore, the purpose of the present study was to investigate a putative mechanism for the age-dependent perception of itch sensation by comparing histamine-induced scratching behaviors in 45-day old (D45) and 75-day old male “young adult” mice. The results indicated that, following histamine administration, the D75 mice spent a longer time scratching than D45 mice. However, the intensity of the calcium influx induced by histamine in primary culture of dorsal root ganglia (DRG) neurons was not different between D45 and D75 mice. Moreover, no apparent difference was observed in mRNA levels of a characteristic His-related receptor and ion channel. Incontrast, the mRNA levels of Toll-Like Receptor 4 (TLR4) were increased approximately by two-fold in D75 DRG compared with D45 DRG. Additionally, D75-derived DRG neurons exhibited enhanced intracellular calcium increase by lipopolysaccharide (LPS, a TLR4 agonist) than those of D45 mice. Furthermore, intensities of calciuminflux induced by histamine were significantly potentiated when co-treated with LPS in D75 DRG neurons, butnot in those of D45 mice. Thus, it appears that D75 mice showed enhanced histamine-induced scratching behaviors not by increased expression levels of histamine-related genes, but probably due to augmented TLR4},

keywords = {Dorsal root ganglia, H1R, Histamine, Itch, Scratching behavior, TLR4, TRPV1},

pubstate = {published},

tppubtype = {article}

}

@article{Jang2017,

title = {Phytotherapeutic effects of the fruits of\textit{Poncirus trifoliata}(L.) Raf. on cancer, inflammation, and digestive dysfunction},

author = {Yongwoo Jang and Eun-Kyung Kim and Won-Sik Shim},

doi = {10.1002/ptr.6008},

issn = {0951-418X},

year  = {2018},

date = {2018-04-00},

journal = {Phytotherapy Research},

volume = {32},

number = {4},

pages = {616--624},

publisher = {Wiley},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jang2018,

title = {Involuntary swimming exercise in pregnant rats disturbs ERK1/2 signaling in embryonic neurons through increased cortisol in the amniotic fluid},

author = {Yongwoo Jang and Byeongjun Lee and Eun-Kyung Kim and Won-Sik Shim and Young Duk Yang and Sung Min Kim},

doi = {10.1016/j.bbrc.2017.11.153},

issn = {0006-291X},

year  = {2018},

date = {2018-01-00},

journal = {Biochemical and Biophysical Research Communications},

volume = {495},

number = {1},

pages = {1208--1213},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2017,

title = {Substitution with a Single Cysteine in the Green Fluorescent Protein-Based Calcium Indicator GCaMP3 Enhances Calcium Sensitivity},

author = {Tae Joon Kim and Ji Young Yoo and Won-Sik Shim},

doi = {10.1007/s10895-017-2159-2},

issn = {1573-4994},

year  = {2017},

date = {2017-11-00},

journal = {J Fluoresc},

volume = {27},

number = {6},

pages = {2187--2193},

publisher = {Springer Science and Business Media LLC},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Afzal2017,

title = {Glucosylsphingosine Activates Serotonin Receptor 2a and 2b: Implication of a Novel Itch Signaling Pathway},

author = {Ramsha Afzal and Won-Sik Shim},

doi = {10.4062/biomolther.2016.207},

issn = {1976-9148},

year  = {2017},

date = {2017-09-01},

journal = {Biomolecules & Therapeutics},

volume = {25},

number = {5},

pages = {497--503},

publisher = {The Korean Society of Applied Pharmacology},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Venkatesan2017,

title = {Lactucopicrin potentiates neuritogenesis and neurotrophic effects by regulating Ca 2+ /CaMKII/ATF1 signaling pathway},

author = {Ramu Venkatesan and Won-Sik Shim and Eui-Ju Yeo and Sun Yeou Kim},

doi = {10.1016/j.jep.2016.12.035},

issn = {0378-8741},

year  = {2017},

date = {2017-02-00},

journal = {Journal of Ethnopharmacology},

volume = {198},

pages = {174--183},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Park2016,

title = {Enhanced Cellular Uptake and Pharmacokinetic Characteristics of Doxorubicin-Valine Amide Prodrug},

author = {Yohan Park and Ju-Hwan Park and Suryeon Park and Song Lee and Kwan Cho and Dae-Duk Kim and Won-Sik Shim and In-Soo Yoon and Hyun-Jong Cho and Han-Joo Maeng},

doi = {10.3390/molecules21101272},

issn = {1420-3049},

year  = {2016},

date = {2016-10-00},

journal = {Molecules},

volume = {21},

number = {10},

publisher = {MDPI AG},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Hwang2015,

title = {A novel synthetic\textit{Piper}amide derivative NED-180 inhibits hyperpigmentation by activating the PI3K and ERK pathways and by regulating Ca^{2+}influx via TRPM1 channels},

author = {Eunson Hwang and Taek Hwan Lee and Wook-Joo Lee and Won-Sik Shim and Eui-Ju Yeo and Sanghee Kim and Sun Yeou Kim},

doi = {10.1111/pcmr.12430},

issn = {1755-1471},

year  = {2016},

date = {2016-01-00},

journal = {Pigment Cell Melanoma Res.},

volume = {29},

number = {1},

pages = {81--91},

publisher = {Wiley},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Pradhananga2015,

title = {Caffeic acid exhibits anti-pruritic effects by inhibition of multiple itch transmission pathways in mice},

author = {Sabindra Pradhananga and Won-Sik Shim},

doi = {10.1016/j.ejphar.2015.06.006},

issn = {0014-2999},

year  = {2015},

date = {2015-09-00},

journal = {European Journal of Pharmacology},

volume = {762},

pages = {313--321},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid26243199,

title = {Amniotic fluid exerts a neurotrophic influence on fetal neurodevelopment via the ERK/GSK-3 pathway},

author = {Yongwoo Jang and Eun-Kyung Kim and Won-Sik Shim and Ki-Min Song and Sung Min Kim},

doi = {10.1186/s40659-015-0029-4},

issn = {0717-6287},

year  = {2015},

date = {2015-08-01},

journal = {Biol Res},

volume = {48},

number = {1},

pages = {44},

abstract = {BACKGROUND: The fetus is surrounded by the amniotic fluid (AF) contained by the amniotic sac of the pregnant female. The AF is directly conveyed to the fetus during pregnancy. Although AF has recently been reported as an untapped resource containing various substances, it remains unclear whether the AF could influence fetal neurodevelopment.nnRESULTS: We used AF that was extracted from embryos at 16 days in pregnant SD rat and exposed the AF to the neural cells derived from the embryos of same rat. We found that the treatment of AF to cortical neurons increased the phosphorylation in ERK1/2 that is necessary for fetal neurodevelopment, which was inhibited by the treatment of MEK inhibitors. Moreover, we found the subsequent inhibition of glycogen synthase kinase-3 (GSK-3), which is an important determinant of cell fate in neural cells. Indeed, AF increased the neural clustering of cortical neurons, which revealed that the clustered cells were proliferating neural progenitor cells. Accordingly, we confirmed the ability of AF to increase the neural progenitor cells through neurosphere formation. Furthermore, we showed that the ERK/GSK-3 pathway was involved in AF-mediated neurosphere enlargement.nnCONCLUSIONS: Although the placenta mainly supplies oxygenated blood, nutrient substances for fetal development, these findings further suggest that circulating-AF into the fetus could affect fetal neurodevelopment via MAP kinases-derived GSK-3 pathway during pregnancy. Moreover, we suggest that AF could be utilized as a valuable resource in the field of regenerative medicine.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jang2015,

title = {Red ginseng extract blocks histamine-dependent itch by inhibition of H1R/TRPV1 pathway in sensory neurons},

author = {Yongwoo Jang and Wook-Joo Lee and Gyu-Sang Hong and Won-Sik Shim},

doi = {10.1016/j.jgr.2015.01.004},

issn = {1226-8453},

year  = {2015},

date = {2015-07-00},

journal = {Journal of Ginseng Research},

volume = {39},

number = {3},

pages = {257--264},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Sundaramoorthy2015,

title = {Modulation of Intracellular Calcium Levels by Calcium Lactate Affects Colon Cancer Cell Motility through Calcium-Dependent Calpain},

author = {Pasupathi Sundaramoorthy and Jae Jun Sim and Yeong-Su Jang and Siddhartha Kumar Mishra and Keun-Yeong Jeong and Poonam Mander and Oh Byung Chul and Won-Sik Shim and Seung Hyun Oh and Ky-Youb Nam and Hwan Mook Kim},

editor = {Neil A. Hotchin},

doi = {10.1371/journal.pone.0116984},

issn = {1932-6203},

year  = {2015},

date = {2015-01-28},

journal = {PLoS ONE},

volume = {10},

number = {1},

publisher = {Public Library of Science (PLoS)},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Maeng2014,

title = {Upregulation of COX-2 in the lung cancer promotes overexpression of multidrug resistance protein 4 (MRP4) via PGE2-dependent pathway},

author = {Han-Joo Maeng and Wook-Joo Lee and Qing-Ri Jin and Ji-Eun Chang and Won-Sik Shim},

doi = {10.1016/j.ejps.2014.05.023},

issn = {0928-0987},

year  = {2014},

date = {2014-10-00},

journal = {European Journal of Pharmaceutical Sciences},

volume = {62},

pages = {189--196},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Maeng2013,

title = {Addition of amino acid moieties to lapatinib increases the anti‐cancer effect via amino acid transporters},

author = {Han‐Joo Maeng and Eun‐Seo Kim and Chieyeon Chough and Misuk Joung and Jee Woong Lim and Chang‐Koo Shim and Won‐Sik Shim},

doi = {10.1002/bdd.1872},

issn = {1099-081X},

year  = {2014},

date = {2014-01-00},

journal = {Biopharm & Drug Disp},

volume = {35},

number = {1},

pages = {60--69},

publisher = {Wiley},

abstract = {ABSTRACTAnti‐cancer agents delivered to cancer cells often show multi‐drug resistance (MDR) due to expulsion of the agents. One way to address this problem is to increase the accumulation of anti‐cancer agents in cells via amino acid transporters. Thus, val‐lapatinib and tyr‐lapatinib were newly synthesized by adding valine and tyrosine moieties, respectively, to the parent anti‐cancer agent lapatinib without stability issues in rat plasma. Val‐lapatinib and tyr‐lapatinib showed enhanced anti‐cancer effects versus the parent lapatinib in various cancer cell lines, including human breast cancer cells (MDA‐MB‐231, MCF7) and lung cancer cells (A549), but not in non‐cancerous MDCK‐II cells. A glutamine uptake study revealed that both val‐lapatinib and tyr‐lapatinib, but not the parent lapatinib, inhibited glutamine transport in MDA‐MB‐231 and MCF7 cells, suggesting the involvement of amino acid transporters. In conclusion, val‐lapatinib and tyr‐lapatinib have enhanced anti‐cancer effects, likely due to an increased uptake of the agents into cancer cells via amino acid transporters. The present data suggest that amino acid transporters may be an effective drug delivery target to increase the uptake of anti‐cancer agents, leading to one method of overcoming MDR in cancer cells. Copyright © 2013 John Wiley & Sons, Ltd.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jang2013,

title = {Naringin Exhibits in vivo Prokinetic Activity via Activation of Ghrelin Receptor in Gastrointestinal Motility Dysfunction Rats},

author = {Yongwoo Jang and Tae-Kwang Kim and Won-Sik Shim},

doi = {10.1159/000354579},

issn = {1423-0313},

year  = {2013},

date = {2013-11-04},

journal = {Pharmacology},

volume = {92},

number = {3-4},

pages = {191--197},

publisher = {S. Karger AG},

abstract = {\textbf{\textit{Background and Purpose:}} Poncirus fructus (PF), also known as the dried immature fruit of \textit{Poncirus trifoliata }(L.) Raf., has long been used as a cure for the treatment of various gastrointestinal disorders in eastern Asia. Recently, it was reported that naringin, a flavonoid constituent of the PF extract, causes the activation of ghrelin receptor in vitro. Although the ghrelin receptor is involved in the enhancement of intestinal motility, there are no studies as yet involving in vivo action of naringin. Therefore, the purpose of the present study is to investigate whether naringin exhibits a prokinetic effect in vivo. \textbf{\textit{Methods:}} We measured the intestinal transit rate in rats with gastrointestinal motility dysfunction (GMD) and performed a pharmacokinetic analysis of naringin to investigate the effect of naringin on prokinetic activity in vivo. \textbf{\textit{Results:}} The results of this study show that the aqueous extract of PF and its constituent naringin have a strong prokinetic activity in GMD rats via activation of the ghrelin receptor. Surprisingly, pharmacokinetic analysis revealed that naringin has low bioavailability (11%), implying that the prokinetic effect of naringin was largely due to the local activation of ghrelin receptor in the intestine rather than a systemic effect after absorption. Indeed, it turned out that intravenous administration of naringin led to a lower prokinetic effect than when administrated orally to rats, indicating that naringin prefers to act on the intestinal wall rather than getting absorbed into the systemic circuit. This local mode of action might be advantageous for preventing possible systemic side effects since naringin is not well absorbed into the system circuit. \textbf{\textit{Conclusions:}} Naringin exhibits an in vivo prokinetic activity by a preferable local activation of ghrelin receptor. Moreover, we propose that naringin could play a role as a leading compound for the development of ghrelin receptor-based prokinetic agents.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid22934579,

title = {An improved prediction of the human in vivo intestinal permeability and BCS class of drugs using the in vitro permeability ratio obtained for rat intestine using an Ussing chamber system},

author = {Hong Li and Hyo-Eon Jin and Won-Sik Shim and Chang-Koo Shim},

doi = {10.3109/03639045.2012.714787},

issn = {1520-5762},

year  = {2013},

date = {2013-10-01},

journal = {Drug Dev Ind Pharm},

volume = {39},

number = {10},

pages = {1515--1522},

abstract = {The Biopharmaceutics Classification System (BCS) was developed to facilitate estimation of the in vivo pharmacokinetic performance of drugs from human intestinal permeability and solubility. However, the measurement of human in vivo intestinal permeability, unlike that of solubility, is problematic and inefficient. Thus, rat in vitro intestinal permeability results obtained via the Ussing chamber technique are often used instead. However, these data could be unreliable due to difficulty in maintaining the viability of the dissected intestinal membrane in the Ussing chamber. Therefore, a more efficient method to obtain a reliable in vitro permeability is mandatory. Here, we propose a new approach by introducing a novel factor called the permeability ratio (PR). Basically, PR is a rat in vitro intestinal permeability obtained from the Ussing chamber, which is then corrected by the permeability of lucifer yellow, a paracellular permeability marker. To prove the validity of the method, 12 model drugs representing different BCS classes were tested, and the correlation with human in vivo intestinal permeability was high. More importantly, the new method perfectly classified all 12 model drugs. The results indicate that PR is a reliable factor with high correlation to human in vivo intestinal permeability, which can further be used to accurately predict the BCS classification.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid22934579b,

title = {An improved prediction of the human in vivo intestinal permeability and BCS class of drugs using the in vitro permeability ratio obtained for rat intestine using an Ussing chamber system},

author = {Hong Li and Hyo-Eon Jin and Won-Sik Shim and Chang-Koo Shim},

doi = {10.3109/03639045.2012.714787},

issn = {1520-5762},

year  = {2013},

date = {2013-10-01},

journal = {Drug Dev Ind Pharm},

volume = {39},

number = {10},

pages = {1515--1522},

abstract = {The Biopharmaceutics Classification System (BCS) was developed to facilitate estimation of the in vivo pharmacokinetic performance of drugs from human intestinal permeability and solubility. However, the measurement of human in vivo intestinal permeability, unlike that of solubility, is problematic and inefficient. Thus, rat in vitro intestinal permeability results obtained via the Ussing chamber technique are often used instead. However, these data could be unreliable due to difficulty in maintaining the viability of the dissected intestinal membrane in the Ussing chamber. Therefore, a more efficient method to obtain a reliable in vitro permeability is mandatory. Here, we propose a new approach by introducing a novel factor called the permeability ratio (PR). Basically, PR is a rat in vitro intestinal permeability obtained from the Ussing chamber, which is then corrected by the permeability of lucifer yellow, a paracellular permeability marker. To prove the validity of the method, 12 model drugs representing different BCS classes were tested, and the correlation with human in vivo intestinal permeability was high. More importantly, the new method perfectly classified all 12 model drugs. The results indicate that PR is a reliable factor with high correlation to human in vivo intestinal permeability, which can further be used to accurately predict the BCS classification.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid24079189,

title = {Smooth muscle relaxation activity of an aqueous extract of dried immature fruit of Poncirus trifoliata (PF-W) on an isolated strip of rat ileum},

author = {Kyu-Sang Kim and Won-Sik Shim and Ike Campomayor dela Peña and Eun-Kyung Seo and Woo-Young Kim and Hyo-Eon Jin and Dae-Duk Kim and Suk-Jae Chung and Jae-Hoon Cheong and Chang-Koo Shim},

issn = {1934-578X},

year  = {2013},

date = {2013-08-01},

journal = {Nat Prod Commun},

volume = {8},

number = {8},

pages = {1143--1148},

abstract = {We demonstrated that an aqueous extract of dried immature fruit of Poncirus trifoliate (PF-W) produces relaxation of intestinal smooth muscle using the ileac strips of a rat. Furthermore, the underlying mechanism of its relaxant activity was investigated. PF-W was prepared using the standard extraction protocol. A 1.5 - 2 cm long rat ileac strip was placed in an organ bath with Tyrode's solution and smooth muscle contractility was recorded by connecting it to a force transducer. Various compounds were added to the organ baths, and changes in muscular contractility were measured. PF-W concentration-dependently induced relaxation of rat ileac strips that were contracted both spontaneously and via acetylcholine treatment. Various potassium channel blockers did not inhibit the relaxation by PF-W. No difference in the effect of PF-W was observed between ileac strips treated with low (20 mM) and high concentrations (60 mM) of KCl. PF-W inhibited the contraction of rat ileac strips induced by extracellular calcium. PF-W acts as a potent smooth muscle relaxant, implicating its possible action as a rapid acting reliever for abdominal pains and a cure for intestinal convulsion. Considering that PF-W also exhibits prokinetic activity, its use in various gastrointestinal disorders seems promising.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jang2013b,

title = {Ghrelin receptor is activated by naringin and naringenin, constituents of a prokinetic agent Poncirus fructus},

author = {Yongwoo Jang and Su-Won Kim and Jungeun Oh and Gyu-Sang Hong and Eun-Kyoung Seo and Uhtaek Oh and Won-Sik Shim},

doi = {10.1016/j.jep.2013.04.039},

issn = {0378-8741},

year  = {2013},

date = {2013-07-00},

journal = {Journal of Ethnopharmacology},

volume = {148},

number = {2},

pages = {459--465},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jin2012,

title = {Transport of gemifloxacin, a 4th generation quinolone antibiotic, in the Caco-2 and engineered MDCKII cells, and potential involvement of efflux transporters in the intestinal absorption of the drug},

author = {Hyo-Eon Jin and Boran Song and Sang-Bum Kim and Won-Sik Shim and Dae-Duk Kim and Saeho Chong and Suk-Jae Chung and Chang-Koo Shim},

doi = {10.3109/00498254.2012.720740},

issn = {1366-5928},

year  = {2013},

date = {2013-04-00},

journal = {Xenobiotica},

volume = {43},

number = {4},

pages = {355--367},

publisher = {Informa UK Limited},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Park2013,

title = {Chitosan microspheres as an alveolar macrophage delivery system of ofloxacin via pulmonary inhalation},

author = {Ju-Hwan Park and Hyo-Eon Jin and Dae-Duk Kim and Suk-Jae Chung and Won-Sik Shim and Chang-Koo Shim},

doi = {10.1016/j.ijpharm.2012.10.044},

issn = {0378-5173},

year  = {2013},

date = {2013-01-00},

journal = {International Journal of Pharmaceutics},

volume = {441},

number = {1-2},

pages = {562--569},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid22941487,

title = {Differential changes in functional activity of organic cation transporters in rats with uranyl nitrate-induced acute renal failure},

author = {Han-Joo Maeng and Won-Sik Shim and Sun-Joo Ahn and Sang-Soo Yu and Dae-Duk Kim and Chang-Koo Shim and Suk-Jae Chung},

doi = {10.1007/s12272-012-0814-3},

issn = {1976-3786},

year  = {2012},

date = {2012-08-01},

journal = {Arch Pharm Res},

volume = {35},

number = {8},

pages = {1441--1448},

abstract = {We studied the impact of experimental kidney failure on the pharmacokinetics of a model organic cation and investigated the underlying mechanism(s) of the organic cation transporters. The systemic pharmacokinetics and tissue distribution of triethylmethylammonium (TEMA), a model organic cation, were characterized after intravenous doses of 0.3-30 μmol/kg in rats with or without uranyl nitrate-induced acute renal failure (UN-ARF). To study the effect of endogenous substrates in plasma from UN-ARF rats on organic cation transport, rOCT- or rOCT2-dependent uptake of tetraethylammonium (TEA) was studied in rOCT1-transfected or rOCT2-transfected LLC-PK1 cells, respectively. As a result, the AUC for TEMA was increased, probably because of decreased total clearance, and the tissue-to-plasma concentration ratio (T/P ratio) of TEMA was unchanged in the liver but decreased significantly in the kidneys of UN-ARF rats. In vitro, the uptake of TEA was decreased significantly by adding UN-ARF plasma, compared with control plasma, in rOCT2-overexpressing LLC-PK1 cells, but not in rOCT1-overexpressing LLC-PK1 cells. These observations suggest that the induction of UN-ARF leads to an accumulation of endogenous organic cation(s), probably rOCT2 substrate(s), in the plasma, thereby affecting the TEMA pharmacokinetics and distribution to the kidneys in rats.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid22233275,

title = {Enhanced intracellular accumulation of a non-nucleoside anti-cancer agent via increased uptake of its valine ester prodrug through amino acid transporters},

author = {Eun-Young Kwak and Won-Sik Shim and Ji-Eun Chang and Saeho Chong and Dae-Duk Kim and Suk-Jae Chung and Chang-Koo Shim},

doi = {10.3109/00498254.2011.646339},

issn = {1366-5928},

year  = {2012},

date = {2012-07-01},

journal = {Xenobiotica},

volume = {42},

number = {7},

pages = {603--613},

abstract = {The phenomenon known as multiple-drug resistance, whereby anti-cancer agents are expelled from cancer cells, makes it necessary to develop methods that will reliably increase the accumulation of anti-cancer agents within cancer cells. To accomplish this goal, a new model compound, Val-SN-38, was synthesized by introducing valine to SN-38, an active ingredient of irinotecan. Val-SN-38 improved intracellular accumulation approximately 5-fold in MCF7 cells, compared with SN-38, and rather than changes in membrane permeability, the amino acid transporter ATB(0,+) played a role, whereas the dipeptide transporter PEPT1 did not. Other sodium-dependent amino acid transporters, namely ATA1, ATA2, and ASCT2, were unexpectedly involved in the uptake of Val-SN-38 as well. The efflux of Val-SN-38 by major efflux transporters was variably changed, but not significantly. In summary, the enhanced accumulation of Val-SN-38 in cancer cells was due to augmented uptake via various amino acid transporters. The results of the present study make a compelling argument in favour of a prodrug concept that can improve intracellular accumulation and take advantage of amino acid transporters without significantly inducing multiple-drug resistance.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid24130931,

title = {Investigation into the Efficacy of Val-SN-38, a Valine-Ester Prodrug of the Anti-Cancer Agent SN-38},

author = {Eun-Young Kwak and Min-Koo Choi and Su-Geun Yang and Chang-Koo Shim and Won-Sik Shim},

doi = {10.4062/biomolther.2012.20.3.326},

issn = {1976-9148},

year  = {2012},

date = {2012-05-01},

journal = {Biomol Ther (Seoul)},

volume = {20},

number = {3},

pages = {326--331},

abstract = {We recently reported that Val-SN-38, a novel valine ester prodrug of SN-38, had greatly improved the intracellular accumulation of SN-38 in MCF-7 cell line, probably through enhanced uptake via amino acid transporters. In the present study, the efficacy of Val-SN-38 was further investigated both in vitro and in vivo. It was found that the in vitro cytotoxic effect of Val-SN-38 was similar to that of SN-38. Moreover, Val-SN-38 exhibited an equal potency to that of SN-38 in survival experiments in vivo. Because these results seemed to be contrary to the previous finding, further investigation was performed to find out the underlying cause of the contradiction. As only the lactone form is known to have cytotoxic activity, the proportion of lactone in Val-SN-38 and SN-38 was determined, but no differences were found. However, it turned out that Val-SN-38 had poor stability compared with SN-38, which resulted in a decrease in beneficial efficacy for Val-SN-38. Overall, the present study showed that a valine-added prodrug approach could be advantageous provided that the stability of the compound can be ensured. We believe this is a noteworthy study that unravels the discrepancy between intracellular accumulation and efficacy of valine-added prodrug.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Chang2011,

title = {Liver Cancer Targeting of Doxorubicin with Reduced Distribution to the Heart Using Hematoporphyrin-Modified Albumin Nanoparticles in Rats},

author = {Ji-Eun Chang and Won-Sik Shim and Su-Geun Yang and Eun-Young Kwak and Saeho Chong and Dae-Duk Kim and Suk-Jae Chung and Chang-Koo Shim},

doi = {10.1007/s11095-011-0603-6},

issn = {1573-904X},

year  = {2012},

date = {2012-03-00},

journal = {Pharm Res},

volume = {29},

number = {3},

pages = {795--805},

publisher = {Springer Science and Business Media LLC},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Yoon2011,

title = {Proliferation and chondrogenic differentiation of human adipose-derived mesenchymal stem cells in porous hyaluronic acid scaffold},

author = {In-Soo Yoon and Chung Wook Chung and Jong-Hyuk Sung and Hyun-Jong Cho and Jung Sun Kim and Won-Sik Shim and Chang-Koo Shim and Suk-Jae Chung and Dae-Duk Kim},

doi = {10.1016/j.jbiosc.2011.06.018},

issn = {1389-1723},

year  = {2011},

date = {2011-10-00},

journal = {Journal of Bioscience and Bioengineering},

volume = {112},

number = {4},

pages = {402--408},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}