Growth Factor/Extracellular Matrix Interactions During Branching Morphogenesis

分支形态发生过程中生长因子/细胞外基质的相互作用

• 批准号: 10931291
• 负责人: Matthew Hoffman
• 金额: $ 74.88万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10931291
• 关键词: Acinar Cell; Adult; Affect; Basement membrane; Binding; Biological; Cell surface; Contracts; Development; Embryonic Development; Enzymes; Epithelial Cells; Epithelium; Epitopes; Extracellular Matrix; FGF10 gene; Family; Fibroblast Growth Factor; Fibroblast Growth Factor Receptors; Frequencies; Genetic; Gland; Growth Factor; Head and Neck Neoplasms; Heparitin Sulfate; Heterogeneity; Human; Impairment; Knock-out; Knockout Mice; Knowledge; Lacrimal gland structure; Measures; Modification; Morphogenesis; Mus; Myoepithelial; Myoepithelial cell; Natural regeneration; Online Mendelian Inheritance In Man; Oral health; Organ; Organ Culture Techniques; Organogenesis; Organoids; Patients; Phenotype; Production; Quality of life; Radiation; Reagent; Role; Saliva; Salivary; Salivary Glands; Signal Transduction; Sjogren' s Syndrome; Specificity; Stains; Sulfate; Syndrome; Therapeutic; Tissue Expansion; Tissues; digital; drinking; drinking behavior; early embryonic stage; fetal; fibroblast growth factor receptor 2b; gland development; insight; novel; progenitor; repaired; response; single-cell RNA sequencing; stem cells; sulfotransferase; tissue regeneration; tool

The exquisite control of growth factor signaling by HS is dictated by the tremendous structural heterogeneity of its sulfated modifications. Heparan sulfotransferase enzymes generate highly 3-O-sulfated epitopes on cell surfaces and in the extracellular matrix. Functional redundancy exists among the family of seven enzymes, but Hs3st3a1 and Hs3st3b1 sulfated HS increases epithelial FGFR signaling and morphogenesis. Single-cell RNAseq analysis of control SMGs identifies increased expression of Hs3st3a1 and Hs3st3b1 in endbud and myoepithelial cells, both of which are progenitor cells during development and regeneration. We previously generated both Hs3st3a1-/- and Hs3st3b1-/- single knockout mice. Salivary glands from both mice have impaired fetal epithelial morphogenesis and reduced 3-O-sulfated HS in the basement membrane. Analysis of adult KO gland function revealed there was an increase in frequency of drinking behavior in both KO mice, suggesting basal salivary hypofunction. We have also generated Hs3st3a1-/- ;Hs3st3b1-/- double knockout (DKO) mice, which are viable, but smaller, with disrupted acinar and myoepithelial cell development. Analysis of HS from the DKO SG shows a loss of specific highly 3-O-sulfated tetrasaccharides. We measured increased increased acinar phospho-Erk staining, with disrupted acinar polarity, suggesting 3-O-sulfated epitopes restrict FGF signaling and their loss increases FGF signaling disrupting acinar differentiation and function. There was also reduced development of myoepithelial cells, which contract to expel saliva. DKO mice have reduced saliva flow and increased drinking frequency. We used novel HS reagents with defined 3-O-sulfated modifications, primary organ cultures and organoids to investigate 3-O-HS-dependent mechanisms regulating acinar and myoepithelial development. We identified that HS-dependent FGFR signaling drives both acinar and MEC development. Thus, loss of s3-O-sulfated epitopes increases FGF signaling, disrupts acinar and MEC development resulting in hypofunction. Understanding how 3-O-sulfation regulates FGFR-dependent myoepithelial progenitor function will be important to manipulate HS-binding growth factors to enhance tissue function and regeneration. This information may be useful to manipulate cellular specificity of HS-binding growth factors and fine-tune biological responses, enhancing progenitor expansion for tissue regeneration. While we focus on the salivary gland, the mouse phenotypes direct us to investigate earlier stages of embryo development and to compare other organs that are affected. These mice are important tools to analyze 3-O-sulfation in salivary gland progenitors and to better understand the fine tuning of cellular responses to FGFRs and HS modifications.

HS对生长因子信号的精致控制取决于其硫酸化修饰的巨大结构异质性。乙酰肝素磺基转移酶在细胞表面和细胞外基质中产生高度3-O的表位。七种酶的家族中存在功能冗余，但是HS3ST3A1和HS3ST3B1硫酸化的HS会增加上皮FGFR信号传导和形态发生。对照SMGS的单细胞RNASEQ分析确定了末端和肌上皮细胞中HS3ST3A1和HS3ST3B1的表达增加，这两个都是在发育和再生过程中的祖细胞。我们以前同时生成了HS3ST3A1 - / - 和HS3ST3B1 - / - 单敲除小鼠。来自两只小鼠的唾液腺都损害了胎儿上皮形态发生，并且在基底膜中降低了3-O的HS。对成年KO腺功能的分析表明，两种KO小鼠的饮用行为频率都有增加，这表明基础唾液功能功能低下。我们还产生了HS3ST3A1 - / - ; HS3ST3B1 - / - 双基因敲除（DKO）小鼠，这些小鼠可行但较小，伴有腺泡和肌上皮细胞的发育。对DKO SG的HS的分析表明，特定高度3-O的四糖的损失。我们测量了增加的增加的磷酸化磷酸化ERK染色，并具有破坏的腺泡极性，这表明3-O阳离子的表位限制了FGF信号，其损耗增加了FGF信号传导破坏腺泡分化和功能。肌上皮细胞的发育也降低，这些细胞会摄入唾液。 DKO小鼠的唾液流量降低并增加了饮酒频率。我们使用具有定义的3-O硫化修饰，原始器官培养物和器官的新型HS试剂来研究调节腺泡和肌上皮发育的3-O-HS依赖性机制。我们确定HS依赖性FGFR信号传导驱动腺泡和MEC发育。因此，S3-O硫化表位的丧失会增加FGF信号传导，破坏腺泡和MEC发育，导致功能障碍。了解3-O硫化如何调节FGFR依赖性肌上皮祖细胞功能对于操纵HS结合生长因子以增强组织功能和再生很重要。该信息可能有助于操纵HS结合生长因子和微调生物学反应的细胞特异性，从而增强祖细胞的扩展以促进组织再生。当我们专注于唾液腺时，小鼠表型指导我们研究胚胎发育的早期阶段，并比较受影响的其他器官。这些小鼠是分析唾液腺祖细胞中3-O硫化的重要工具，并更好地了解细胞对FGFR和HS修饰的细胞反应的微调。

项目成果

O-glycosylation modulates integrin and FGF signalling by influencing the secretion of basement membrane components.

O-糖基化通过影响基底膜成分的分泌来调节整合素和 FGF 信号传导。

• DOI: 10.1038/ncomms1874
• 发表时间: 2012-05-29
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Tian E;Hoffman MP;Ten Hagen KG
• 通讯作者: Ten Hagen KG

Anatomy, biogenesis and regeneration of salivary glands.

• DOI: 10.1159/000358776
• 发表时间: 2014
• 期刊: Monographs in oral science
• 作者: Holmberg KV;Hoffman MP
• 通讯作者: Hoffman MP

Luminal signaling: it's what's on the inside that counts.

发光信号：内部的东西才是最重要的。

• DOI: 10.1016/j.devcel.2014.11.019
• 发表时间: 2014
• 期刊: Developmental cell
• 影响因子: 11.8
• 作者: Symonds,JenniferM;Hoffman,MatthewP
• 通讯作者: Hoffman,MatthewP

Salivary gland development: a template for regeneration.

• DOI: 10.1016/j.semcdb.2013.12.001
• 发表时间: 2014-01
• 期刊: SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY
• 影响因子: 7.3
• 作者: Patel, Vaishali N.;Hoffman, Matthew P.
• 通讯作者: Hoffman, Matthew P.

Loss of Hs3st3a1 or Hs3st3b1 enzymes alters heparan sulfate to reduce epithelial morphogenesis and adult salivary gland function.

• DOI: 10.1016/j.matbio.2021.10.002
• 发表时间: 2021-09
• 期刊: Matrix biology : journal of the International Society for Matrix Biology
• 作者: Patel VN;Pineda DL;Berenstein E;Hauser BR;Choi S;Prochazkova M;Zheng C;Goldsmith CM;van Kuppevelt TH;Kulkarni A;Song Y;Linhardt RJ;Chibly AM;Hoffman MP
• 通讯作者: Hoffman MP