Impact of N-Acetylgalactosamine-4-Sulfatase (Arylsulfatase B) on the Progression of Melanoma

N-乙酰半乳糖胺-4-硫酸酯酶（芳基硫酸酯酶 B）对黑色素瘤进展的影响

• 批准号: 10258903
• 负责人: Joanne Kramer Tobacman
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10258903
• 关键词: Address; Affect; Apoptosis; Arylsulfatase B; Binding; Biological; Breast; CD34 gene; Cardiac; Cell Line; Cell Survival; Cell physiology; Cells; Characteristics; Chondroitin; Chondroitin Sulfate A; Chondroitin Sulfate Proteoglycan; Clinical; Coculture Techniques; Colon; Dermatan Sulfate; Disease; Doctor of Medicine; Doctor of Philosophy; Dose; Early Diagnosis; Effectiveness; Enzymes; Event; Excision; Family; Family member; Galectin 3; Gelatinase A; General Population; Genetic Transcription; Growth; Human; Immune; Immune checkpoint inhibitor; Immunooncology; Implant; Insulin Receptor; Investigation; Knockout Mice; Lead; Liver; Lymphocyte; Lysosomal Storage Diseases; MAP Kinase Gene; MAPK3 gene; MAPK8 gene; MMP2 gene; Malignant Neoplasms; Matrix Metalloproteinases; Measures; Mediator of activation protein; Melanoma Cell; Metastatic Melanoma; Modification; Morbidity - disease rate; Mucopolysaccharidosis VI; Mus; Mutation; Neurologic; Nuclear Translocation; Oncologist; Orthopedics; PTPN11 gene; Pathology; Patients; Phosphorylation; Prostate; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Publications; Publishing; Radial; Recombinants; Recurrence; Reporting; Research Personnel; Role; Signal Transduction; Signaling Molecule; Stage at Diagnosis; Structural defect; Sulfatases; Sulfate; Testing; Time; Tissue Microarray; Tissues; Toxic effect; Transcription Coactivator; Treatment Effectiveness; Treatment Protocols; Veterans; Work; Xenograft Model; anti-PD1 therapy; base; cancer cell; checkpoint inhibition; checkpoint therapy; chondroitin sulfate glycosaminoglycan; enzyme activity; experience; experimental study; human tissue; humanized mouse; immunomodulatory therapies; insight; interest; knock-down; lead sulfate; melanocyte; melanoma; mortality; mouse model; novel; novel strategies; novel therapeutic intervention; overexpression; p38 Mitogen Activated Protein Kinase; pembrolizumab; polysulfated glycosaminoglycan; programmed cell death ligand 1; receptor binding; respiratory; response; transcription factor; transcriptome sequencing

The morbidity and mortality of malignant melanoma are significant problems for veterans, their families, and the general public. Progress has been made in early diagnosis and in treatment, in particular with advances in checkpoint inhibition therapy. However, not all patients respond to this approach and the treatment has major toxicity. There is a continuing unmet need for improvement in treatment of melanoma. This project presents a novel and potentially breakthrough treatment approach, based on the impact of the enzyme N- acetylgalactosamine-4-sulfatase, also known as arylsulfatase B (ARSB). This enzyme removes sulfate groups from chondroitin 4-sulfate (C4S) and dermatan sulfate and is required for the degradation of these sulfated glycosaminoglycans. Our previous work has shown that lower ARSB activity is associated with more aggressive melanomas. Also, decline in ARSB leads to increased expression of the melanoma proteoglycan chondroitin sulfate proteoglycan (CSPG)4 and of the matrix metalloproteinase pro-MMP2 which facilitates invasion. Other experiments have shown increase in PD-L1 expression in melanoma cells following ARSB silencing. In this project, we will determine the transcriptional mechanism by which decline in ARSB increases expression of PD-L1 in normal melanocytes and melanoma cells. Experiments will show how decline and increase in ARSB affect melanoma cell survival and intracellular signaling. The impact of anti-PD1 treatment on melanoma cell survival will be tested in live cell co-culture with immune cells following ARSB silencing. Other studies in the B16F10 and YUMM mouse models of melanoma and in a humanized mouse xenograft model will show the impact of modulation of ARSB in association with checkpoint inhibitor treatment on the progression of primary and metastatic melanomas. This project is based on over 30 publications in which Dr. Tobacman and collaborators have identified biological consequences of decline in ARSB. A previous report with project collaborator, Arkadiusz Dudek, M.D., Ph.D., a distinguished oncologist and investigator with strong interest, background, and clinical experience in immuno-oncology, identified decline in ARSB with increasing aggressiveness of melanoma cell lines, as well as significant increases in expression CSPG4 and pro-MMP2. Inborn deficiency of ARSB is present in the lysosomal storage disease Mucopolysaccharidosis (MPS) VI, in which mutations lead to marked reduction of ARSB activity. In malignant cells from prostate, breast, colon, and liver, as well as in melanoma cells and tissue, we have shown that expression and activity of ARSB are reduced, compared to normal melanocytes and tissue. Decline in ARSB leads to accumulation of the sulfated glycosaminoglycans chondroitin 4-sulfate and dermatan sulfate, from which ARSB normally removes the 4- sulfate group at the non-reducing end and is required to initiate their degradation. With decline in ARSB, C4S accumulates and its interactions with critical molecules are disrupted. We have shown that galectin-3, a co- transcriptional activator which binds to other important molecules, including the insulin receptor, binds less to more highly sulfated C4S and has increased nuclear translocation and interaction with transcription factors. Inversely, SHP2 (PTPN11), the ubiquitous non-membrane Src homology region 2 (SH2)-containing protein tyrosine phosphatase 2, binds more with more highly sulfated C4S and is less available for dephosphorylation of critical signaling molecules, including phospho-ERK1/2. Hence, the transcriptional events arising from altered sulfation of C4S have profound impact on vital cellular processes. This project will provide new insight into how changes in ARSB, and the resulting changes in chondroitin 4-sulfation and signaling and transcriptional events, impact on melanoma progression and on response to checkpoint inhibition. The findings may lead to new approaches to treatment of melanoma and to improvement in response to checkpoint inhibition, resulting in reduced suffering, morbidity, and mortality from melanoma.

恶性黑色素瘤的发病率和死亡率对于退伍军人及其家人来说是一个重大问题。 和广大公众。在早期诊断和治疗方面取得了进展，特别是在 在检查点抑制疗法中。然而，并非所有患者都对这种方法有反应，并且治疗有很大的影响。 毒性。对改进黑色素瘤治疗的需求一直未得到满足。该项目提出了一个 基于酶 N- 的影响，新颖且具有潜在突破性的治疗方法 乙酰半乳糖胺-4-硫酸酯酶，也称为芳基硫酸酯酶 B (ARSB)。该酶可去除硫酸根基团 来自 4-硫酸软骨素 (C4S) 和硫酸皮肤素，是降解这些硫酸盐所必需的 糖胺聚糖。我们之前的工作表明，较低的 ARSB 活性与更具攻击性相关 黑色素瘤。此外，ARSB 的下降会导致黑色素瘤蛋白聚糖软骨素的表达增加 硫酸盐蛋白聚糖 (CSPG)4 和促进侵袭的基质金属蛋白酶原 MMP2。其他 实验表明，ARSB 沉默后，黑色素瘤细胞中 PD-L1 的表达增加。 在这个项目中，我们将确定 ARSB 下降增加的转录机制 PD-L1在正常黑色素细胞和黑色素瘤细胞中的表达。实验将显示如何衰退和 ARSB 的增加影响黑色素瘤细胞的存活和细胞内信号传导。抗PD1治疗的影响 ARSB 沉默后，将在活细胞与免疫细胞共培养中测试黑色素瘤细胞的存活率。其他 B16F10 和 YUMM 黑色素瘤小鼠模型以及人源化小鼠异种移植模型的研究将 显示与检查点抑制剂治疗相关的 ARSB 调节对疾病进展的影响 原发性和转移性黑色素瘤。该项目基于 30 多份出版物，其中 Tobacman 博士和 合作者已经确定了 ARSB 下降的生物学后果。之前的项目报告 合作者，Arkadiusz Dudek，医学博士、哲学博士，一位杰出的肿瘤学家和研究者，有着浓厚的兴趣， 免疫肿瘤学的背景和临床经验表明，ARSB 随着增加而下降 黑色素瘤细胞系的侵袭性，以及 CSPG4 和 pro-MMP2 表达的显着增加。 先天性 ARSB 缺陷存在于溶酶体贮积病粘多糖贮积症 (MPS) VI 中， 其中突变导致 ARSB 活性显着降低。在前列腺、乳腺、结肠的恶性细胞中， 和肝脏，以及黑色素瘤细胞和组织中，我们已经证明 ARSB 的表达和活性 与正常黑素细胞和组织相比减少。 ARSB 的下降导致硫酸盐的积累 糖胺聚糖软骨素 4-硫酸盐和硫酸皮肤素，ARSB 通常从中去除 4- 硫酸基团位于非还原端，是启动其降解所必需的。随着 ARSB、C4S 的下降 积累并且其与关键分子的相互作用被破坏。我们已经证明半乳糖凝集素 3 (galectin-3) 与其他重要分子（包括胰岛素受体）结合的转录激活剂，与 C4S 硫酸化程度更高，并增加了核易位以及与转录因子的相互作用。 相反，SHP2 (PTPN11)，普遍存在的含有非膜 Src 同源区 2 (SH2) 的蛋白质 酪氨酸磷酸酶 2，与高度硫酸化的 C4S 结合更多，并且不太可用于去磷酸化 关键信号分子，包括磷酸化 ERK1/2。因此，由改变引起的转录事件 C4S 的硫酸化对重要的细胞过程具有深远的影响。 该项目将为 ARSB 的变化以及软骨素的变化提供新的见解 4-硫酸化、信号传导和转录事件，对黑色素瘤进展和反应的影响 检查点抑制。这些发现可能会带来治疗黑色素瘤和改善黑色素瘤的新方法 响应检查点抑制，从而减少黑色素瘤的痛苦、发病率和死亡率。