Cardiotrophin-Like Cytokine 1, a Candidate Molecule for the FSGS Factor

心肌营养素样细胞因子 1,FSGS 因子的候选分子

基本信息

项目摘要

DESCRIPTION (provided by applicant): Steroid resistant nephrotic syndrome and focal segmental glomerulosclerosis (FSGS) pose especially difficult clinical problems because the disease recurs in about 30% of such patients after renal transplantation and leads to transplant loss in many patients. The molecular basis for recurrent FSGS is unknown although abnormal podocyte proteins associated with familial FSGS have been identified. We have documented that serum or plasma from FSGS patients increases glomerular albumin permeability (Palb) and that the specific activity of certain protein fractions of FSGS plasma is more than 10,000-fold greater than that of the initial plasma. Intravenous injection of this fraction causes proteinuria in rats. We have recently used state-of-the-art proteomics techniques to study the proteins in the active plasma fraction from a FSGS patient. We identified a single cytokine, cardiotrophin-like cytokine 1 (CLC-1). CLC-1 is a candidate for the FSGS permeability factor because of this unique presence in active fractions of FSGS plasma. CLC-1 is also overexpressed in peripheral blood cells of patients with recurrent FSGS. CLC-1 is a cytokine of the IL-6 family that is synthesized by cells in many tissues. CLC-1 rapidly increases Palb in a concentration-dependent fashion. The magnitude and time-course of this increase are comparable to that induced by FSGS plasma. The permeability-effect of FSGS plasma as well as of recombinant CLC-1 itself is blocked by anti-CLC-1 antibody. We have found that receptor proteins for CLC-1 are present on glomeruli. The proposed studies are based on a.) the novel discovery of CLC-1 in the active fraction of FSGS plasma, b.) the overexpression CLC-1 in blood cells of patients with FSGS, c.) the expression of receptor proteins in glomeruli, d.) the vigorous activity of CLC-1 in the Palb assay and finally, e.) the striking ability of anti-CLC-1 monoclonal antibodies to block the permeability effect of FSGS plasma. Our central hypothesis is that CLC-1 is the elusive FSGS permeability factor. We will pursue the following Specific Aims. SA1: Confirm the relevance of CLC-1 to activity of the FSGS factor. We will test the sera of 4 patients with FSGS for the presence of CLC-1 using LC-MS/MS and Western blotting for CLC-1 using polyclonal antibodies. Patients with FSGS and rapid progression and/or prior post-transplant recurrence will be studied. Plasma from normal persons and from non-FSGS patients undergoing plasmapheresis prior to renal transplantation will be used as a controls. We will confirm that CLC-1 accounts for permeability activity by showing that antibodies to CLC-1 block permeability activity of FSGS plasma fractions of at least 10 patients. SA2: Confirm that CLC-1 increases permeability and study the mechanism by which this occurs. We will define the effect of CLC-1 on permeability using concentrations of CLC-1 from 0.05 to 50 ng/ml in established Palb and GEC monolayer protocols. Specificity of effects will be tested using blocking antibodies to CLA-1 and known blockers of FSGS plasma effects. Preliminary studies show that CLC-1 increases glomerular permeability in a concentration dependent manner and that anti- CLC-1 blocks this effect. We will determine if the addition of CLC-1 binding-partners cytokine-like factor-1 (CLF-1) or soluble ciliary neurotrophic factor receptor (sCNTFR) enhances the CLC-1 permeability effect. We will study the expression of each of the components of the receptors for CLC- 1 (gp 130, CNTF receptor-1 (CNTFR-1), and leukemia inhibitory factor receptor (LIFR)) by glomeruli and cultured using RT-PCR and Western blotting. These components will be localized in glomeruli and cultured GEC using immunohistochemistry. Preliminary studies show that anti-CLC-1 blocks the effect of FSGS serum and that glomeruli express RNA for receptor components. These studies will verify that CLC-1 is a candidate for the "FSGS permeability factor" and will identify the potential receptor responsible for its permeability effect on GEC. Future studies will permit characterization of the signaling pathways that control the permeability barrier and design of specific and effective therapies for proteinuria in FSGS, and potentially, for proteinuria in other renal diseases. PUBLIC HEALTH RELEVANCE: Kidney disease that is identified by the appearance of large amounts of protein in the urine, edema or swelling of tissues, low plasma proteins and increased cholesterol is termed nephrotic syndrome. In many cases, nephrotic syndrome can be treated effectively with the use of artificial steroid hormones such as prednisone. In some patients with nephrotic syndrome that does not respond to treatment, microscopic examination of kidney tissue shows patchy or segmental scarring of some of the filters, which are called glomeruli. This pattern of damage is called focal segmental glomerulosclerosis or FSGS. Glomeruli are responsible for making urine and for preventing protein leak. When they are damaged, protein leaks into the urine and, if damage and protein leak continue, further scarring of the entire kidney leads to kidney failure. FSGS a leading cause of kidney failure in children and causes kidney failure in many adults, as well. Genetic abnormalities are present in some children and adults with FSGS, but there is no known genetic basis for FSGS in most patients. FSGS recurs in some patients with FSGS who have kidney failure and receive kidney transplants. The recurrence appears to be related to a small protein in the plasma that injures the glomeruli and leads to protein leak and scarring. The nature and the source of this protein are not known. During the past 20 years, we have studied serum samples from more than 500 patients with FSGS. We have used a variety of biochemical techniques for identifying proteins and determining their characteristics. These techniques include gel chromatography to determine the protein size and electrical charge, affinity chromatography to determine binding properties, and mass spectrometry. We have recently identified a protein, cardiotrophin-like cytokine 1 (CLC- 1), as a candidate for the active substance. This protein is best known for its role in nerve development and maturation. It is also important in the development of the kidney. We have tested this substance and found that even very brief exposure of glomeruli leads to severe increase in permeability in a manner identical to the reaction to FSGS proteins. We have also shown that blood cells of affected patients make more than normal amounts of CLC-1, and that antibodies to CLC-1 prevent the injury caused by patients' plasma. Glomeruli have the proteins that bind to CLC-1 and permit it to act on cells. The presence of CLC-1 in the plasma of a patient with FSGS, its overexpression by patients' blood cells, its activity in injuring glomeruli, and that fact that antibody to CLC blocks the effect of FSGS plasma lead us to the proposed studies to define its role in this serious kidney disease. We will perform studies to verify the presence of CLC-1 in samples from several patients with FSGS and recurrence after transplantation. We will use mass spectrometry and other advanced techniques of protein analysis for these studies. We will also test the effect of purified CLC-1 on glomeruli and on glomerular cells in culture. The results will verify our preliminary finding that CLC-1 is a component of the plasma of FSGS patients and that it injures glomeruli in a manner that is comparable to the injury caused by FSGS proteins. We will do further studies to identify the cell receptor responsible for CLC-1 activity on the glomerulus and to determine the cell responses that are required for its actions. These studies will point to new ways to treat FSGS and to prevent it from progressing to kidney failure or from recurring in newly transplanted kidneys.
描述(由申请人提供):类固醇抵抗性肾病综合征和局灶节段性肾小球硬化(FSGS)是特别困难的临床问题,因为约30%的此类患者在肾移植后会复发,并导致许多患者移植失败。虽然已经发现了与家族性FSGS相关的异常足细胞蛋白,但复发性FSGS的分子基础尚不清楚。我们已经证实,FSGS患者的血清或血浆会增加肾小球白蛋白通透性(Palb),并且FSGS血浆中某些蛋白质组分的比活性比初始血浆高出10,000倍以上。静脉注射该组分可引起大鼠蛋白尿。我们最近使用了最先进的蛋白质组学技术来研究FSGS患者活性血浆部分中的蛋白质。我们鉴定了一种单一的细胞因子,心促因子样细胞因子1 (CLC-1)。CLC-1是FSGS渗透因子的候选物,因为它独特地存在于FSGS血浆的活性组分中。CLC-1在复发性FSGS患者的外周血中也过表达。CLC-1是一种IL-6家族的细胞因子,由许多组织的细胞合成。CLC-1以浓度依赖的方式迅速增加Palb。这种增加的幅度和时间过程与FSGS等离子体诱导的相当。FSGS血浆的透性作用以及重组CLC-1本身被抗CLC-1抗体阻断。我们发现肾小球上存在CLC-1受体蛋白。提出的研究是基于a)在FSGS血浆活性部分中发现CLC-1的新发现,b) FSGS患者血细胞中CLC-1的过表达,c)肾小球中受体蛋白的表达,d) Palb试验中CLC-1的强烈活性,最后,e)抗CLC-1单克隆抗体阻断FSGS血浆渗透作用的惊人能力。我们的中心假设是CLC-1是难以捉摸的FSGS渗透因子。具体目标是:SA1:确认CLC-1与FSGS因子活性的相关性。我们将使用LC-MS/MS和Western blotting检测4例FSGS患者的血清中是否存在CLC-1,并使用多克隆抗体检测CLC-1。FSGS快速进展和/或移植后复发的患者将被研究。正常人和肾移植前行血浆置换的非fsgs患者的血浆作为对照。我们将通过显示CLC-1抗体阻断至少10例患者FSGS血浆组分的渗透性活性来证实CLC-1对渗透性活性的影响。SA2:证实CLC-1增加渗透率,并研究其发生的机制。我们将在已建立的Palb和GEC单层方案中使用0.05至50 ng/ml浓度的CLC-1来确定CLC-1对渗透率的影响。将使用CLA-1的阻断抗体和已知的FSGS血浆效应阻滞剂来测试效果的特异性。初步研究表明,CLC-1以浓度依赖的方式增加肾小球通透性,而抗CLC-1可阻断这一作用。我们将确定CLC-1结合伙伴细胞因子样因子-1 (CLF-1)或可溶性纤毛神经营养因子受体(sCNTFR)的加入是否增强了CLC-1的通透性作用。我们将使用RT-PCR和Western blotting技术研究肾小球中CLC- 1受体(gp130、CNTF受体-1 (CNTFR-1)和白血病抑制因子受体(LIFR))各组分的表达。这些成分将定位在肾小球和培养GEC使用免疫组织化学。初步研究表明,抗clc -1阻断了FSGS血清的作用,肾小球表达RNA受体成分。这些研究将验证CLC-1是“FSGS渗透性因子”的候选者,并将确定其对GEC渗透性影响的潜在受体。未来的研究将允许表征控制渗透性屏障的信号通路和设计特异性和有效的治疗FSGS蛋白尿的方法,并可能用于其他肾脏疾病的蛋白尿。公共卫生相关性:肾脏疾病,通过尿液中出现大量蛋白质,组织水肿或肿胀,血浆蛋白低和胆固醇升高,被称为肾病综合征。在许多情况下,使用人工类固醇激素如强的松可以有效地治疗肾病综合征。在一些治疗无效的肾病综合征患者中,肾组织的显微镜检查显示一些滤过物出现斑片状或节段性瘢痕,这些滤过物被称为肾小球。这种损害模式被称为局灶节段性肾小球硬化或FSGS。肾小球负责制造尿液和防止蛋白质泄漏。当它们受损时,蛋白质会渗漏到尿液中,如果损害和蛋白质泄漏继续下去,整个肾脏的进一步疤痕会导致肾衰竭。FSGS是儿童肾功能衰竭的主要原因,也会导致许多成年人肾功能衰竭。一些患有FSGS的儿童和成人存在遗传异常,但大多数患者的FSGS没有已知的遗传基础。一些肾功能衰竭并接受肾移植的FSGS患者会复发。复发似乎与血浆中一种小蛋白质损伤肾小球并导致蛋白质泄漏和瘢痕形成有关。这种蛋白质的性质和来源尚不清楚。在过去的20年里,我们研究了500多名FSGS患者的血清样本。我们已经使用了多种生物化学技术来鉴定蛋白质并确定它们的特性。这些技术包括测定蛋白质大小和电荷的凝胶色谱法、测定结合性质的亲和色谱法和质谱法。我们最近确定了一种蛋白质,心促因子样细胞因子1 (CLC- 1),作为活性物质的候选。这种蛋白质以其在神经发育和成熟中的作用而闻名。它在肾脏的发育中也很重要。我们对这种物质进行了测试,发现即使非常短暂地暴露在肾小球中,也会导致通透性的严重增加,其反应方式与对FSGS蛋白的反应相同。我们还表明,受影响的患者的血细胞产生的CLC-1多于正常水平,并且针对CLC-1的抗体可以防止患者血浆造成的损伤。肾小球中含有与CLC-1结合并允许其作用于细胞的蛋白质。CLC-1在FSGS患者血浆中的存在,其在患者血细胞中的过度表达,其在肾小球损伤中的活性,以及CLC抗体阻断FSGS血浆作用的事实,导致我们提出研究以确定其在这种严重肾脏疾病中的作用。我们将进行研究,以验证CLC-1在几例移植后复发的FSGS患者样本中的存在。我们将使用质谱和其他先进的蛋白质分析技术进行这些研究。我们还将测试纯化的CLC-1对肾小球和培养的肾小球细胞的影响。该结果将验证我们的初步发现,即CLC-1是FSGS患者血浆中的一种成分,并且其损伤肾小球的方式与FSGS蛋白引起的损伤相当。我们将做进一步的研究,以确定负责肾小球上CLC-1活性的细胞受体,并确定其作用所需的细胞反应。这些研究将指出治疗FSGS的新方法,并防止其发展为肾衰竭或在新移植的肾脏中复发。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Virginia J. Savin其他文献

Virginia J. Savin的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Virginia J. Savin', 18)}}的其他基金

Cytokine based murine focal glomerulosclerosis model a prelude to novel therapy
基于细胞因子的鼠局灶性肾小球硬化模型是新疗法的前奏
  • 批准号:
    8696811
  • 财政年份:
    2011
  • 资助金额:
    $ 16.21万
  • 项目类别:
Cytokine based murine focal glomerulosclerosis model a prelude to novel therapy
基于细胞因子的鼠局灶性肾小球硬化模型是新疗法的前奏
  • 批准号:
    8143226
  • 财政年份:
    2011
  • 资助金额:
    $ 16.21万
  • 项目类别:
Cytokine based murine focal glomerulosclerosis model a prelude to novel therapy
基于细胞因子的鼠局灶性肾小球硬化模型是新疗法的前奏
  • 批准号:
    8255321
  • 财政年份:
    2011
  • 资助金额:
    $ 16.21万
  • 项目类别:
Cytokine based murine focal glomerulosclerosis model a prelude to novel therapy
基于细胞因子的鼠局灶性肾小球硬化模型是新疗法的前奏
  • 批准号:
    8398954
  • 财政年份:
    2011
  • 资助金额:
    $ 16.21万
  • 项目类别:
Cardiotrophin-Like Cytokine 1, a Candidate Molecule for the FSGS Factor
心肌营养素样细胞因子 1,FSGS 因子的候选分子
  • 批准号:
    7988466
  • 财政年份:
    2009
  • 资助金额:
    $ 16.21万
  • 项目类别:
Cardiotrophin-Like Cytokine 1, a Candidate Molecule for the FSGS Factor
心肌营养素样细胞因子 1,FSGS 因子的候选分子
  • 批准号:
    7588208
  • 财政年份:
    2009
  • 资助金额:
    $ 16.21万
  • 项目类别:
ETIOLOGIC FACTORS IN FOCAL GLOMERULAR SCLEROSIS
局灶性肾小球硬化的病因
  • 批准号:
    3245224
  • 财政年份:
    1993
  • 资助金额:
    $ 16.21万
  • 项目类别:
ETIOLOGIC FACTOR IN FOCAL GLOMERULOSCLEROSIS
局灶性肾小球硬化症的病因
  • 批准号:
    2693159
  • 财政年份:
    1993
  • 资助金额:
    $ 16.21万
  • 项目类别:
ETIOLOGIC FACTORS IN FOCAL GLOMERULAR SCLEROSIS
局灶性肾小球硬化的病因
  • 批准号:
    2143254
  • 财政年份:
    1993
  • 资助金额:
    $ 16.21万
  • 项目类别:
ETIOLOGIC FACTORS IN FOCAL GLOMERULAR SCLEROSIS
局灶性肾小球硬化的病因
  • 批准号:
    2143253
  • 财政年份:
    1993
  • 资助金额:
    $ 16.21万
  • 项目类别:

相似海外基金

Co-designing a lifestyle, stop-vaping intervention for ex-smoking, adult vapers (CLOVER study)
为戒烟的成年电子烟使用者共同设计生活方式、戒烟干预措施(CLOVER 研究)
  • 批准号:
    MR/Z503605/1
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Research Grant
Early Life Antecedents Predicting Adult Daily Affective Reactivity to Stress
早期生活经历预测成人对压力的日常情感反应
  • 批准号:
    2336167
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Standard Grant
RAPID: Affective Mechanisms of Adjustment in Diverse Emerging Adult Student Communities Before, During, and Beyond the COVID-19 Pandemic
RAPID:COVID-19 大流行之前、期间和之后不同新兴成人学生社区的情感调整机制
  • 批准号:
    2402691
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Standard Grant
Elucidation of Adult Newt Cells Regulating the ZRS enhancer during Limb Regeneration
阐明成体蝾螈细胞在肢体再生过程中调节 ZRS 增强子
  • 批准号:
    24K12150
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Migrant Youth and the Sociolegal Construction of Child and Adult Categories
流动青年与儿童和成人类别的社会法律建构
  • 批准号:
    2341428
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Standard Grant
Understanding how platelets mediate new neuron formation in the adult brain
了解血小板如何介导成人大脑中新神经元的形成
  • 批准号:
    DE240100561
  • 财政年份:
    2024
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Discovery Early Career Researcher Award
Laboratory testing and development of a new adult ankle splint
新型成人踝关节夹板的实验室测试和开发
  • 批准号:
    10065645
  • 财政年份:
    2023
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Collaborative R&D
Usefulness of a question prompt sheet for onco-fertility in adolescent and young adult patients under 25 years old.
问题提示表对于 25 岁以下青少年和年轻成年患者的肿瘤生育力的有用性。
  • 批准号:
    23K09542
  • 财政年份:
    2023
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Identification of new specific molecules associated with right ventricular dysfunction in adult patients with congenital heart disease
鉴定与成年先天性心脏病患者右心室功能障碍相关的新特异性分子
  • 批准号:
    23K07552
  • 财政年份:
    2023
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Issue identifications and model developments in transitional care for patients with adult congenital heart disease.
成人先天性心脏病患者过渡护理的问题识别和模型开发。
  • 批准号:
    23K07559
  • 财政年份:
    2023
  • 资助金额:
    $ 16.21万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了