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，他们的家人来说是重大问题 和公众。在早期诊断和治疗方面已经取得了进展，尤其是随着进步的进展 在检查点抑制疗法中。但是，并非所有患者都对此方法做出反应，并且治疗的主要 毒性。持续未满足的黑色素瘤治疗需求。这个项目提出了 基于酶N-的影响，新型和潜在的突破治疗方法 乙酰乳糖胺-4-硫酸酯酶，也称为芳基硫酸酯酶B（ARSB）。该酶去除了硫酸盐基团 从软骨素4-硫酸盐（C4S）和硫酸皮肤硫酸盐，是这些硫酸盐降解所必需的 糖胺聚糖。我们以前的工作表明，较低的ARSB活动与更具侵略性有关 黑色素瘤。同样，ARSB的下降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和基质金属蛋白酶pro-MMP2的硫酸盐蛋白聚糖（CSPG），可促进入侵。其他 实验表明，ARSB沉默后黑色素瘤细胞中PD-L1表达的增加。 在这个项目中，我们将确定ARSB下降的转录机制 PD-L1在正常黑色素细胞和黑色素瘤细胞中的表达。实验将显示下降和 ARSB的增加会影响黑色素瘤细胞的存活和细胞内信号传导。抗PD1处理对 黑色素瘤细胞存活率将在ARSB沉默后与免疫细胞共培养中测试。其他 在B16F10和Yumm小鼠模型和人源化小鼠异种移植模型中的研究将 显示ARSB调制与检查点抑制剂治疗对进展的影响 原发性和转移性黑色素瘤。该项目基于托巴克曼博士和 合作者确定了ARSB下降的生物学后果。项目的先前报告 合作者，Arkadiusz Dudek，M.D。 背景和免疫肿瘤学方面的临床经验确定ARSB的下降随着增加 黑色素瘤细胞系的侵略性，以及表达CSPG4和Pro-MMP2的显着增加。 ARSB的先天缺乏存在于溶酶体储存疾病粘二糖（MPS）VI中， 其中突变导致ARSB活性显着降低。在前列腺，乳房，结肠的恶性细胞中， 肝脏以及黑色素瘤细胞和组织中，我们已经表明ARSB的表达和活性是 与正常的黑素细胞和组织相比，减少了。 ARSB的下降导致硫化的积累 糖胺聚糖4-硫酸软骨素和硫酸皮肤素，ARSB通常从中去除4-- 在非还原端处的硫酸盐组，必须启动其降解。随着ARSB的下降，C4S 累积及其与关键分子的相互作用受到破坏。我们已经证明了Galectin-3，一个共同 与其他重要分子结合的转录激活剂，包括胰岛素受体，与 更高硫酸化的C4，并增加了核易位和与转录因子的相互作用。 相反，SHP2（PTPN11），无处不在的非膜SRC同源性区域2（SH2） - 含蛋白 酪氨酸磷酸酶2与更高硫酸化的C4结合，较少可用于去磷酸化 临界信号分子，包括磷酸-ERK1/2。因此，因改变而产生的转录事件 C4的硫酸化对重要的细胞过程有深远的影响。 该项目将为ARSB的变化以及软骨素的变化提供新的见解 4-硫化，信号传导和转录事件，对黑色素瘤进展的影响以及对 检查点抑制。这些发现可能会导致黑色素瘤治疗和改善的新方法 响应检查点抑制，导致黑色素瘤的痛苦，发病率和死亡率降低。