Tyrosinase-related Protein Research Articles

Abstract LB233: Single-cell transcriptional analysis of embryonic melanoblasts pinpoints embryonic cell states that underlie targeted therapy and immune checkpoint blockade resistance

Abstract Melanoma is a malignant neoplasia of pigment-producing melanocytes notable for its ability to metastasize early during primary tumor growth. Melanomas display typically high mutation burden—a hallmark associated with immune checkpoint blockade therapy (ICBT) sensitivity—but can still evade ICBT. These melanoma phenotypes likely relate to embryonic neural crest cells, a population of cells from which melanocytes arise. For instance, BRAF inhibition-evading melanoma cells adopt neural crest-like states. However, the exact embryonic lineage underlying BRAFi resistance remains unknown as does its relationship with any neural crest-like states that underlie ICBT resistance. To discover such states, we used a mouse fluorescent reporter model of melanoblast development in which melanocyte-specific GFP is driven by the dopachrome tautomerase promoter (iDct-GFP). Embryonic day (E) 11.5 and 15.5 melanoblasts were isolated and sequenced using single-cell transcriptomics (scRNA-seq). These time-points enriched for cells migrating from the dermis to epidermis (at E11.5) or for cells colonizing the hair follicle (at E15.5). These characteristics could help metastatic progression, namely migration across the basement membrane in early invasive melanomas and colonization of metastatic sites. We classified these embryonic melanocytes and their precursors into multiple clusters which in turn were grouped into four key lineages: Neural Progenitor, Schwann Cell Progenitor (SCP), Mesencyhmal (Mes) and Melanocytes (Mel). To detect the presence of these states in tumors, we derived developmental gene modules (DGMs): sets of genes that were preferentially co-expressed in each embryonic cell state. DGMs stratified TCGA samples; SCP-high and Mes-high subtypes were associated with worse survival compared to Mel-high tumors. Moreover, SCP-high tumors had lower T cell infiltration, and were associated with higher NGFR expression, a known marker of ICBT resistance. High expression of SCP DGMs was associated with poor response to anti-PD1 ICBT. Targeting factors underlying the SCP state might therefore sensitize tumours to ICBT. Mes DGMs on the other hand were associated with targeted therapy resistance. Cross-referencing DGMs with published RNA-seq data from melanoma cell lines suggest that SCP and Mes DGMs are induced by pro-inflammatory cytokines secreted by immune cells in the tumor microenvironment. Together, these observations suggest that the SCP state is implicated in the phenomenon of inflammation-induced de-differentiation, a process by which melanomas are able to evade ICBT. Our identification of the SCP and Mes states, along with a deeper understanding of their transcriptional programs and signaling pathways, may reveal clinically actionable targets to disrupt immuno- or targeted-therapy resistance. Citation Format: Vishaka Gopalan, Chun Wai Wong, Chi-Ping Day, Eva Perez-Guijarro, Maxwell P. Lee, Yuhong Jiang, Howard H. Yang, Maira Alves-Constantino, Antonella Sassano, Cari Smith, Mark Simpson, Sung Chin, Jessica Ebersole, Charli Gruen, Emily Wu, Sridhar Hannenhalli, Adam Hurlstone, Glenn Merlino, Kerrie Marie. Single-cell transcriptional analysis of embryonic melanoblasts pinpoints embryonic cell states that underlie targeted therapy and immune checkpoint blockade resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB233.

Gene expression profiles of skin from cyclin dependent kinases 5-knockdown mice

Objective: This study aimed to identify genes regulated by cyclin dependent kinases 5 (CDK5) that participate in hair pigmentation in mice.Methods: The mRNA expression profiles of skin samples from <i>CDK5</i>-knockdown mice were constructed using high-throughput RNA sequencing and compared with those of wild-type mice.Results: In total, 8,002 known genes were differentially expressed between <i>CDK5</i>-knockdown and wild-type mice. Of these, 3,658 were upregulated and 4,344 were downregulated in the skin of <i>CDK5</i>-knockdown mice. An additional 318 previously unknown genes were also differentially expressed, with 171 downregulated and 147 upregulated genes in the skin of <i>CDK5</i>-knockdown mice. Of the known genes expressed in mouse skin, 80 were associated with hair color, with 61 showing lower expression and 19 exhibiting higher expression in skin of <i>CDK5</i>-knockdown mice. Importantly, the expression of the tyrosinase-related protein 1 (<i>TYRP1</i>) and the calcium signaling pathway were also found to be regulated by CDK5, suggesting that pigmentation is regulated by CDK5 via the calcium signaling pathway and TYRP1.Conclusion: The transcriptome profiles obtained from the skin of <i>CDK5</i>-knockdown mice compared to wild-type mice provide a valuable resource to help understand the mechanism by which CDK5 regulates melanogenesis in mice and other animals.

In silico molecular docking of luteolin as a potential antihyperpigmentation agent

Excessive melanin synthesis, often triggered by overexposure to UV rays, is catalyzed by melanogenesis enzymes such as tyrosinase, tyrosinase-related protein 1, and D-dopachrome tautomerase. Derived from natural sources, the flavonoid compound luteolin is explored for its antihyperpigmentation potential. This study assesses luteolin’s efficacy as an antihyperpigmentation agent by analyzing its affinity and bond interactions with melanogenesis enzymes through an in silico approach. Molecular docking, facilitated by HyperChem 8 for test compound optimization and Chimera 1.11.1 for protein preparation, alongside method validation and docking with AutoDockTools 1.5.6, established the protocol’s validity with an RMSD value of ≤3 Å. Docking results reveal luteolin's higher affinity for the target proteins compared to native ligands, with binding energies of -5.63 kcal/mol for tyrosinase, -6.18 kcal/mol for tyrosinase-related protein 1, and -6.54 kcal/mol for D-dopachrome tautomerase. The interaction between luteolin and these proteins involves hydrogen, hydrophobic, electrostatic, and Van der Waals bonds, with amino acid residues His61, Lys129, Arg132 (tyrosinase); His192, His224, Val89 (tyrosinase-related protein 1); and Ile64, Asn73 (D-dopachrome tautomerase) participating in hydrogen bond formation. These findings suggest luteolin’s significant potential as an antihyperpigmentation agent by inhibiting melanogenesis enzymes.

Abstract 6748: Intramuscular delivery of tumor associated antigen DNA vaccines elicits strong cellular immune response, delays tumor growth and prolongs survival

Abstract The development of effective cancer vaccines that can trigger strong and enduring memory T cell responses against specific tumor associated antigens (TAAs) is crucial for advancing immunotherapy. DNA-based cancer vaccines have shown promise in generating precise and long-lasting immune responses. In particular DNA vaccines are capable of inducing both humoral and cellular immune responses. In this study, we aimed to investigate the effectiveness of DNA vaccines targeting the mouse Tyrosine-related protein-2 (Trp2) and human New York esophageal squamous cell carcinoma 1 (NY-ESO-1) TAAs in mouse melanoma model. These DNA vaccines were formulated with a functionalized polymer designed to protect DNA from degradation and improve its bioavailability. DNA vaccines targeting both mouse Trp2 (pVACTR) and human NYESO-1 (pVACNY) were constructed. A mouse melanoma cell line (B16F10-NY) overexpressing human NYESO-1 was also generated and demonstrated to induce tumors in C57BL/6 mice. Female C57BL/6 mice received two doses of intramuscular (i.m.) injection of formulated pVACTR or pVACNY or a combination thereof, formulated with the synthetic functionalized polymer, with a 3-week interval. The DNA vaccination induced a potent T cell response, as evidenced by ELISPOT assays showing strong IFN-γ secreting T cells against NYESO-1 and Trp2. Flow cytometry analysis of spleenocytes showed strong IFN-γ and TNF-α secreting CD4 T cells against NYESO-1. We further evaluated the prophylactic and therapeutic efficacy of DNA vaccines targeting both Trp2 and NYESO-1 using a murine syngeneic model. Animals were challenged with a lethal dose of B16F10-NY two weeks after the second immunization exhibited a significant (p<0.0001) delay in tumor progression and a substantial increase in survival compared to the mock control group. In the therapeutic testing, animals challenged with B16F10-NY and subsequently vaccinated at 4, 11, and 25 days also demonstrated delayed tumor growth and prolonged survival. These findings indicate that formulated DNA vaccine delivered via i.m. triggers robust T cell responses and exhibit both prophylactic and therapeutic efficacy, leading to significantly prolonged survival and delayed tumor growth. A DNA-based vaccine independent of viral vector or device has potential to elicit potent anti-tumor responses with better safety and compliance. Citation Format: Kempaiah Rayavara, John Henderson, Meredyth Kinsella, Jessica Kim, Majed Matar, Subeena Sood, Olivia Signer, Chelsey Bellmon, Corinne Le Goff, Khursheed Anwer, Jean Boyer. Intramuscular delivery of tumor associated antigen DNA vaccines elicits strong cellular immune response, delays tumor growth and prolongs survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6748.

Phase 1, first-in-human study of TYRP1-TCB (RO7293583), a novel TYRP1-targeting CD3 T-cell engager, in metastatic melanoma: active drug monitoring to assess the impact of immune response on drug exposure.

Although checkpoint inhibitors (CPIs) have improved outcomes for patients with metastatic melanoma, those progressing on CPIs have limited therapeutic options. To address this unmet need and overcome CPI resistance mechanisms, novel immunotherapies, such as T-cell engaging agents, are being developed. The use of these agents has sometimes been limited by the immune response mounted against them in the form of anti-drug antibodies (ADAs), which is challenging to predict preclinically and can lead to neutralization of the drug and loss of efficacy. TYRP1-TCB (RO7293583; RG6232) is a T-cell engaging bispecific (TCB) antibody that targets tyrosinase-related protein 1 (TYRP1), which is expressed in many melanomas, thereby directing T cells to kill TYRP1-expressing tumor cells. Preclinical studies show TYRP1-TCB to have potent anti-tumor activity. This first-in-human (FIH) phase 1 dose-escalation study characterized the safety, tolerability, maximum tolerated dose/optimal biological dose, and pharmacokinetics (PK) of TYRP1-TCB in patients with metastatic melanoma (NCT04551352). Twenty participants with cutaneous, uveal, or mucosal TYRP1-positive melanoma received TYRP1-TCB in escalating doses (0.045 to 0.4 mg). All participants experienced ≥1 treatment-related adverse event (TRAE); two participants experienced grade 3 TRAEs. The most common toxicities were grade 1-2 cytokine release syndrome (CRS) and rash. Fractionated dosing mitigated CRS and was associated with lower levels of interleukin-6 and tumor necrosis factor-alpha. Measurement of active drug (dual TYPR1- and CD3-binding) PK rapidly identified loss of active drug exposure in all participants treated with 0.4 mg in a flat dosing schedule for ≥3 cycles. Loss of exposure was associated with development of ADAs towards both the TYRP1 and CD3 domains. A total drug PK assay, measuring free and ADA-bound forms, demonstrated that TYRP1-TCB-ADA immune complexes were present in participant samples, but showed no drug activity in vitro. This study provides important insights into how the use of active drug PK assays, coupled with mechanistic follow-up, can inform and enable ongoing benefit/risk assessment for individuals participating in FIH dose-escalation trials. Translational studies that lead to a better understanding of the underlying biology of cognate T- and B-cell interactions, ultimately resulting in ADA development to novel biotherapeutics, are needed.

Genomic insights into shank and eggshell color in Italian local chickens

Eggshell and shank color in poultry is an intriguing topic of research due to the roles in selection, breed recognition, and environmental adaptation. This study delves into the genomics foundations of shank and eggshell pigmentation in Italian local chickens through genome-wide association studies analysis to uncover the mechanisms governing these phenotypes. To this purpose, 483 animals from 20 local breeds (n = 466) and 2 commercial lines (n = 17) were considered and evaluated for shank and eggshell color. All animals were genotyped using the Affymetrix Axiom 600 K Chicken Genotyping Array. As regards shank color, the most interesting locus was detected on chromosome Z, close to the TYRP1 gene, known to play a key role in avian pigmentation. Additionally, several novel loci and genes associated with shank pigmentation, skin pigmentation, UV protection, and melanocyte regulation were identified (e.g., MTAP, CDKN2A, CDKN2B). In eggshell, fewer significant loci were identified, including SLC7A11 and MITF on chromosomes 4 and 12, respectively, associated with melanocyte processes and pigment synthesis. This comprehensive study shed light on the genetic architecture underlying shank and eggshell color in Italian native chicken breeds, contributing to a better understanding of this phenomenon which plays a role in breed identification and conservation, and has ecological and economic implications.

설견초와 병풀 추출물의 미백활성

This study attempted to assess the whitening activities of Salvia plebeia R. Br. and Centella asiatica extracts which were provided by MEIDEME CO., LTD., their safety was confirmed through cell viability testing. The whitening activities were evaluated by inhibition of melanin synthesis and reduction of whitening-related transcription factors and protein production. The results found that no statistically significant cytotoxicity was observed at 10㎍/㎖, 20㎍/㎖and 40㎍/㎖. In addition, melanin production decreased by 28.4% at Salvia plebeia R. BR and Centella asiatica extract(SCE) 40㎍/㎖. In transcription factors, tyrosinase, tyrosianse related protein(TRP-1) and dopachrome tautomerase(TRP-2) decreased by 17%, 21% and 22% respectively at SCE 40㎍/㎖, confirming statistical significance. Protein production also showed statistically significant results with decrease in tyrosinase, TRP-1 and TRP-2 by 87%, 40% and 27% each at SCE 40㎍/㎖. The above results confirm that SCE is effective in promoting whitening and would be available as a cosmetic ingredient for whitening cosmetics.

Bioactive Fractions Isolated from Harungana madagascariensis Lam. and Psorospermum aurantiacum Engl. Regulate Collagen and Melanin Biosynthesis Gene Expression in UVB-Irradiated Cells with Additional Anti-Inflammatory Potential.

Harungana madagascariensis (HM) and Psorospermum aurantiacum (PA), used traditionally for skin care, have been reported to upregulate the expression of intracellular antioxidant genes, thereby preventing melanoma and protecting fibroblast cell lines from Ultraviolet B (UVB)-induced intracellular oxidative stress. This investigation aimed to identify major compounds in bioactive fractions using bioassay- guided fractionation. The anti-inflammatory effect of fractions was determined by measuring their inhibitory activity on 15-lipoxygenase and nitric oxide (NO) in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. Additionally, the anti-aging efficacy of the fractions was determined by assessing the expression of markers for the aging process, i.e., expression of tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), procollagen type-1 (COL1A1), and matrix metalloproteinase- 1 (MMP-1) in UVB-induced photoaging in skin cell-lines. Furthermore, UHPLCMS- based identification of the bioactive compounds from the most prominent fraction was also carried out. Hexane fraction of HM significantly inhibited (p <0.05) the 15-lipoxygenase (IC50 = 46.80 μg/mL) and NO production (IC50 = 66.55 μg/mL), whereas hexane fraction of PA was effective (p <0.05) in inhibiting 15-lipoxygenase activity (IC50 = 27.55 μg/mL). Furthermore, the hexane fraction of HM and methanol fraction of PA were significantly effective (p <0.05) in reverting the UVB-mediated altered expressions of MMP-1, TYR, TRP-1, and COL1A1. Furthermore, hexane fraction of HM revealed the presence of harunganin and betulinic acid, whereas vismion D, vismin, kenganthranol B, and bianthrone 1a were identified from the methanol fraction of PA. Overall, the hexane fraction of HM and methanol fraction of PA displayed effective anti-aging activities, with additional anti-inflammatory effects.

Effects of Aspirin Eugenol Ester on Liver Oxidative Damage and Energy Metabolism in Immune-Stressed Broilers.

The aim of this study was to investigate the effects of aspirin eugenol ester (AEE) on liver oxidative damage and energy metabolism in immune-stressed broilers. In total, 312 broilers were divided into 4 groups (saline, LPS, SAEE, and LAEE). Broilers in the saline and LPS groups were fed a basal diet; the SAEE and LAEE groups had an added 0.01% AEE in their diet. Broilers in the LPS and LAEE groups were injected with lipopolysaccharides, while the saline and SAEE groups were injected with saline. Results showed that AEE increased the body weight, average daily gain, and average daily feed intake, as well as decreasing the feed conversion ratio of immune-stressed broilers. AEE protects against oxidative damage in immune-stressed broiler livers by elevating the total antioxidant capacity, superoxide dismutase activity, and glutathione S-transferase alpha 3 (GSTA3) and glutaredoxin 2 (GLRX2) expression, while decreasing malondialdehyde content. AEE lessened inflammation by reducing prostaglandin-F2α production and prostaglandin-endoperoxide synthase 2 (PTGS2) and interleukin-1beta (IL-1β) expression. AEE decreased oxidative phosphorylation rates by increasing succinic acid levels and lowering both adenosine diphosphate (ADP) levels and ceroid lipofuscinosis neuronal 5 (CLN5) expression. AEE modulated the metabolism of phenylalanine, tyrosine, lipids, and cholesterol by reducing the phenyllactate and L-arogenate levels, lowering dopachrome tautomerase (DCT) and apolipoprotein A4 (APOA4) expression, and increasing phenylpyruvic acid and dopa decarboxylase (DDC) expression. In summary, AEE can effectively alleviate liver oxidative damage and energy metabolism disorders in immune-stressed broilers.

Patchouli Alcohol: A Potent Tyrosinase Inhibitor Derived from Patchouli Essential Oil with Potential in the Development of a Skin-Lightening Agent

The inhibitory effects of Pogostemon cablin essential oil (patchouli essential oil, PEO) and its primary bioactive compound, patchouli alcohol (PA), on tyrosinase and melanin were investigated in vitro and ex vivo. Treatment with PEO and PA significantly, as well as dose-dependently, reduced forskolin (FRK)-induced melanin biosynthesis, cellular tyrosinase activity, and tyrosinase (TYR) protein expression. However, the transcriptional levels of TYR and tyrosinase-related proteins (TRP-1 and TRP-2) remained unaffected. These results suggest that PEO and PA may directly interrupt tyrosinase enzyme activity, leading to a reduction in melanin biosynthesis. Further experiments supported this notion, revealing that both PEO and PA significantly and dose-dependently inhibited mushroom tyrosinase activity in both the monophenolase and diphenolase phases. Additionally, an in silico molecular docking analysis was performed, utilizing a homology model of human tyrosinase. In conclusion, these findings strongly suggest that patchouli essential oil and its primary bioactive component, patchouli alcohol, hold promise as potential treatments for hyperpigmentary skin conditions and in the development of cosmetic products designed to lighten the skin.

Effect of Petai Bark Extract Gel on Tyrosinase and TRP1 Gene Expression (In Vivo Experimental Study on Hyperpigmented Wistar Mice Exposed to UVB Model)

UVB irradiation can induce the formation of Reactive Oxygen Species (ROS) which causes activation of melanin synthesis through activation of tyrosinase and tyrosinase related protein-1 (TRP1). Secondary metabolites contained in petai peel extract are known to play a role in inhibiting ROS production due to exposure to UVB rays. This study aims to determine the effect of administering petai peel extract gel on the expression of the tyrosinase and TRp1 genes in mouse skin tissue exposed to UVB. The research design was posttest only control group with a completely randomized design method. The samples studied were 24 mice exposed to UVB light with a wavelength of 302 nm and an energy of 390mJ/cm2/day 3 times a week for 2 weeks. This research was carried out in four groups, namely the healthy group (K1), the negative control group (K2), treatment 1 (K3) with 10% petai peel extract gel and treatment 2 (K4) with 20% petai peel extract gel. Tyrosinase and TRP1 gene expression was analyzed using qRT-PCR. qRT-PCR analysis showed that there was a significant decrease in tyrosinase and TRP1 gene expression between groups K3 (tyrosinase 3,19±2,12 and TRP1 4,96±3,42) and K4 (tyrosinase 0,65±0,44 and TRP1 2,22±1,18) compared to group K2 (tyrosinase 17,92±3,77 and TRP1 35,91±4,52). Administration of petai peel extract gel can reduce the expression of tyrosinase and TRP1 genes in hyperpigmentation mice exposed to UVB light.

Exploring the potential of white birch sap: A natural alternative to traditional skin whitening agents with reduced side effects

Common tyrosinase (TYR) inhibitors used in cosmetics, such as hydroquinone, kojic acid, and arbutin, can cause side effects including erythema, skin peeling, and dryness. Therefore, the development of natural whitening agents that offer excellent permeability, minimal irritation, and high safety has become a primary focus in the field of TYR inhibitors. In this study, we demonstrate that White birch sap (WBS), within a safe concentration range, effectively reduces TYR activity and melanin content in both B16F10 mouse melanoma cells and zebrafish larvae. Importantly, WBS exhibits minimal irritation to neutrophils in fluorescent zebrafish and does not affect the behavior of adult zebrafish. Furthermore, WBS downregulates the gene expression levels of microphthalmia-associated transcription factor, TYR, tyrosinase-related protein-1, and tyrosinase-related protein-2 in B16F10 cells. In conclusion, our research confirms that WBS, a naturally derived substance, offers high safety and mild effects, making it a promising candidate for a skin-whitening agent.

Premature gray hair development in the interbrow region owing to the loss of maxillary first molars in young mice.

The exact sites of premature hair graying and whether tooth loss causes this condition remain unknown. In this study, we aimed to explore the effect of reduced mastication on premature hair graying. Maxillary first molars were extracted from young mice, and the mice were observed for 3 months, along with non-extraction control group mice. After 3 months, gray hair emerged in the interbrow region of mice in the tooth extraction group but not in the control group. The expression of tyrosinase-related protein-2 (TRP-2) mRNA was lower in the interbrow tissues of young mice without maxillary molars than in those with maxillary molars. Tooth loss leads to interbrow gray hair growth, possibly because of weakened trigeminal nerve input, suggesting that reduced mastication causes premature graying. Thus, prompt prosthetic treatment after molar loss is highly recommended.

Optimization, Metabolomic Analysis, Antioxidant Potential andDepigmenting Activity of Polyphenolic Compounds fromUnmature Ajwa Date Seeds (Phoenix dactylifera L.) Using Ultrasonic-Assisted Extraction.

This study sought to optimize the ultrasonic-assisted extraction of polyphenolic compounds from unmature Ajwa date seeds (UMS), conduct untargeted metabolite identification and assess antioxidant and depigmenting activities. Response surface methodology (RSM) utilizing the Box-Behnken design (BBD) and artificial neural network (ANN) modeling was applied to optimize extraction conditions, including the ethanol concentration, extraction temperature and time. The determined optimal conditions comprised the ethanol concentration (62.00%), extraction time (29.00 min), and extraction temperature (50 °C). Under these conditions, UMS exhibited total phenolic content (TPC) and total flavonoid content (TFC) values of 77.52 ± 1.55 mgGAE/g and 58.85 ± 1.12 mgCE/g, respectively, with low relative standard deviation (RSD%) and relative standard error (RSE%). High-resolution mass spectrometry analysis unveiled the presence of 104 secondary metabolites in UMS, encompassing phenols, flavonoids, sesquiterpenoids, lignans and fatty acids. Furthermore, UMS demonstrated robust antioxidant activities in various cell-free antioxidant assays, implicating engagement in both hydrogen atom transfer and single electron transfer mechanisms. Additionally, UMS effectively mitigated tert-butyl hydroperoxide (t-BHP)-induced cellular reactive oxygen species (ROS) generation in a concentration-dependent manner. Crucially, UMS showcased the ability to activate mitogen-activated protein kinases (MAPKs) and suppress key proteins including tyrosinase (Tyr), tyrosinase-related protein-1 and -2 (Trp-1 and -2) and microphthalmia-associated transcription factor (MITF), which associated melanin production in MNT-1 cell. In summary, this study not only optimized the extraction process for polyphenolic compounds from UMS but also elucidated its diverse secondary metabolite profile. The observed antioxidant and depigmenting activities underscore the promising applications of UMS in skincare formulations and pharmaceutical developments.

CAR-T cell therapy targeting surface expression of TYRP1 to treat cutaneous and rare melanoma subtypes

A major limitation to developing chimeric antigen receptor (CAR)-T cell therapies for solid tumors is identifying surface proteins highly expressed in tumors but not in normal tissues. Here, we identify Tyrosinase Related Protein 1 (TYRP1) as a CAR-T cell therapy target to treat patients with cutaneous and rare melanoma subtypes unresponsive to immune checkpoint blockade. TYRP1 is primarily located intracellularly in the melanosomes, with a small fraction being trafficked to the cell surface via vesicular transport. We develop a highly sensitive CAR-T cell therapy that detects surface TYRP1 in tumor cells with high TYRP1 overexpression and presents antitumor activity in vitro and in vivo in murine and patient-derived cutaneous, acral and uveal melanoma models. Furthermore, no systemic or off-tumor severe toxicities are observed in an immunocompetent murine model. The efficacy and safety profile of the TYRP1 CAR-T cell therapy supports the ongoing preparation of a phase I clinical trial.

Evaluation and identification of green pharmacophore as a potent inhibitor of human tyrosinase related protein 1 using molecular drug interaction, drug likeness and ADMET analyses

Evaluation and identification of green pharmacophore as a potent inhibitor of human tyrosinase related protein 1 using molecular drug interaction, drug likeness and ADMET analyses

The Antimelanogenic Activity of the Extract of Heukharang Lettuce (Lactuca sativa L.) Leaf

Heukharang (Lactuca sativa L.), the artificially developed lettuce cultivar for enhancement of lactucin content, exhibits antioxidant activities and sleep-promoting effects. However, potential of Heukharang as a raw material for the skin-whitening agent has not been investigated yet. This study evaluated the effects of Heukharang extract (HHE) on α-melanocyte-stimulating hormone (α-MSH)-induced melanogenesis in vitro and in vivo models. Our findings revealed that HHE (25–100 μg/mL) effectively inhibited α-MSH-induced melanin synthesis in both 2D and 3D cell culture environments without compromising the viability of the B16F10 murine melanoma cell line. Furthermore, HHE suppressed intracellular tyrosinase activity, along with both gene and protein expression of tyrosinase (TYR) and tyrosinase-related protein (TRP)-1 and TRP-2. HHE downregulated the mRNA level of microphthalmia-associated transcription factor (MITF) and the phosphorylation of cAMP response element-binding (CREB) protein. The antimelanogenic activity of HHE (50 and 100 μg/mL) was also confirmed using a zebrafish embryo model, highlighting its efficacy in inhibiting α-MSH-induced melanogenesis through the downregulating CREB/MITF/TYR signaling pathways. Moreover, our study demonstrated that a corresponding amount of lactucin (5–20 μM) to the HHE inhibited and regulated melanogenesis. These results collectively suggest that HHE, enriched with lactucin, holds promise as a potential skin-whitening agent for nutricosmetic industries.

Methylanthranilate, a Food Fragrance Attenuates Skin Pigmentation through Downregulation of Melanogenic Enzymes by cAMP Suppression.

Methyl anthranilate (MA) is a botanical fragrance used in food flavoring with unexplored potential in anti-pigment cosmetics. MA dose-dependently reduced melanin content without affecting cell viability, inhibited dendrite elongation and melanosome transfer in the co-culture system of human melanoma cells (MNT-1) and human keratinocyte cell line (HaCaT), and downregulated melanogenic genes, including tyrosinase, tyrosinase-related protein 1 and 2 (TRP-1, TRP-2). Additionally, MA decreased cyclic adenosine monophosphate (cAMP) production and exhibited a significant anti-pigmentary effect in Melanoderm™. These results suggest that MA is a promising anti-pigmentary agent for replacing or complementing existing anti-pigmentary cosmetics.

Melanin Inhibitory Effect of Tuber himalayense Isolated in Incheon, Korea.

There has been a growing interest in skin beauty and antimelanogenic products. Melanogenesis is the process of melanin synthesis whereby melanocytes are activated by UV light or hormone stimulation to produce melanin. Melanogenesis is mediated by several enzymes, such as tyrosinase (TYR), microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), and TRP-2. In this study, we investigated the effect of Tuber himalayense extract on melanin synthesis in α-melanocyte-stimulating hormone (α-MSH)-treated B16F10 melanoma cells. We confirmed that T. himalayense extract was not toxic to α-MSH-treated B16F10 melanoma cells and exhibited a significant inhibitory effect on melanin synthesis at concentrations of 25, 50, and 100 μg/ml. Additionally, the T. himalayense extract inhibited melanin, TRP-1, TRP-2, tyrosinase, and MITF, which are enzymes involved in melanin synthesis, in a concentration-dependent manner. Furthermore, T. himalayense extract inhibited the mitogen-activated protein kinase (MAPK) pathways, such as extracellular signal-regulated kinase-1/2 (ERK), c-Jun N-terminal kinase (JNK), and p38. Therefore, we hypothesized that various components of T. himalayense extract affect multiple factors involved in melanogenesis in B16F10 cells. Our results indicate that T. himalayense extract could potentially be used as a new material for preparing whitening cosmetics.

(2-Methylbutyryl)shikonin Naturally Occurring Shikonin Derivative Ameliorates the α-MSH-Induced Melanogenesis via ERK1/2 and p38 MAP Kinase-Mediated Down-Regulation of the MITF Transcription Factor.

Cutaneous pigmentation is an important phenotypic trait whose regulation, despite recent advances, has yet to be completely elucidated. Melanogenesis, a physiological process of melanin production, is imperative for organism survival as it provides protection against the environmental insults that majorly involve sunlight-induced skin photodamage. However, immoderate melanin synthesis can cause pigmentation disorders associated with a psychosocial impact. In this study, the hypopigmentation effect of (2-methylbutyryl)shikonin, a natural product present in the root extract of Lithospermum erythrorhizon, and the underlying mechanisms responsible for the inhibition of melanin synthesis in α-MSH-stimulated B16F10 cells and C57BL/6J mice was studied. Non-cytotoxic concentrations of (2-methylbutyryl)shikonin significantly repressed cellular tyrosinase activity and melanin synthesis in both in vitro and in vivo models (C57BL/6J mice). (2-Methylbutyryl)shikonin remarkably abolished the protein expression of MITF, tyrosinase, tyrosinase-related protein 1, and tyrosinase-related protein 2, thereby blocking the production of pigment melanin via modulating the phosphorylation status of MAPK proteins, viz., ERK1/2 and p38. In addition, specific inhibition of ERK1/2 attenuated the inhibitory effects of (2-methylbutyryl)shikonin on melanin synthesis, whereas selective inhibition of p38 augmented the inhibitory effect of BSHK on melanin synthesis. Moreover, topical application of (2-methylbutyryl)shikonin on C57BL/6J mouse tails remarkably induced tail depigmentation. In conclusion, with these findings, we, for the first time, report the hypopigmentation effect of (2-methylbutyryl)shikonin via inhibition of cellular tyrosinase enzyme activity, subsequently ameliorating the melanin production, thereby indicating that (2-methylbutyryl)shikonin is a potential natural therapy for hyperpigmentation disorders.