Osteoskeletal regenerative ability of exosomes derived from adipose-derived stem cells upon inhibition of Transforming Growth Factor-β-signaling

抑制转化生长因子-β-信号传导后脂肪干细胞衍生的外泌体的骨骨骼再生能力

基本信息

项目摘要

Tumors, trauma or surgery can lead to critical-sized bone defects requiring extensive reconstruction. Autologous or allogenic transplantation or prosthetic material for bone reconstruction comes with considerable drawbacks and other options need to be explored. Transforming growth factor beta (TGFβ) has been identified to play an important role in bone metabolism. It is hypothesized that inhibition of TGFβ signaling promotes bone regeneration. Small molecule SB431542, a TGFβ inhibitor, leads to increased osteogenic differentiation, thus presenting a valid approach in alternate treatment of bone defects. Tissue engineering using culture-expanded mesenchymal stem cells (MSC) has been shown to be highly effective in experimental research of bone regeneration. However, MSC engraftment and differentiation at the injury site has been shown to be poor. It has been proposed that MSCs secrete bioactive factors, which act directly by promoting intracellular signaling or indirectly by stimulating neighboring cells. Exosomes released from MSCs are biomolecular nanostructures delivering miRNA, proteins and other molecules.We hypothesize that exosomes act as vehicles for autocrine and/or paracrine signaling factors emanating from human adipose tissue-derived stem cells (ex-hASCs) and can thus be isolated from the cells and applied directly to the bone defect. We believe that pretreatment with SB431542 enhances the osteoskeletal repair ability of derived exosomes. Our proposed project aims at investigating the bone regeneration capacity of ex-hASCs treated with SB431542, in comparison to ex-hASCs isolated from untreated hASCs. Moreover, we will characterize the cargo content of ex-hASCs with the goal to identify specific factors playing a key role in enhancing the osteoskelatal regeneration of calvarial bones.First we will isolate the exosomes from untreated and SB431542 treated hASCs. Through in vitro experiments we will analyse the osteoinductive activity of ex-hASCs upon inhibition of TGFβ signaling, using RT-PCR, qPCR and immunoblotting. Osteoskeletal regenerative capacity of ex-hASCs will be analysed in a calvarial defect model using micro-CT, histology and immunofluorescence staining. The cargo content of the isolated exosomes will be characterised via imaging techniques, immunoblotting, proteomic analysis and miRNA analysis upon inhibition of TGFβ-signaling versus exosomes from untreated hASCs. Lastly we will perform functional testing of candidate proteins and/or miRNAs that we earlier identified using gain- and loss-of-function experiments. We believe that exosomes can be used as vehicles for targeted drug delivery and can be preconditioned genetically to improve therapy at the target site. Thus, exosomes could potentially replace cell-based therapy and eradicate the risk of neoplastic transformation of MSC derived cell transplantation.
肿瘤、创伤或手术可能导致严重的骨缺损,需要广泛的重建。自体或异体移植或假体材料用于骨重建有相当大的缺点,需要探索其他选择。转化生长因子β (TGFβ)在骨代谢中起重要作用。据推测,抑制tgf - β信号传导可促进骨再生。小分子SB431542是一种TGFβ抑制剂,可导致成骨分化增加,因此为骨缺损的替代治疗提供了一种有效的方法。利用培养扩增间充质干细胞(MSC)进行组织工程在骨再生的实验研究中已被证明是非常有效的。然而,干细胞在损伤部位的植入和分化表现不佳。有人提出,间充质干细胞分泌生物活性因子,其作用直接通过促进细胞内信号传导或间接通过刺激邻近细胞。间充质干细胞释放的外泌体是一种生物分子纳米结构,可传递miRNA、蛋白质和其他分子。我们假设外泌体作为自分泌和/或旁分泌信号因子的载体,来自人类脂肪组织来源的干细胞(前hascs),因此可以从细胞中分离出来并直接应用于骨缺损。我们认为SB431542预处理可以增强衍生外泌体的骨修复能力。我们提出的项目旨在研究SB431542处理的前hASCs的骨再生能力,与未处理的hASCs分离的前hASCs进行比较。此外,我们将表征前hascs的货物含量,目的是确定在增强颅骨骨骨再生中起关键作用的特定因素。首先,我们将从未经处理和SB431542处理的hASCs中分离外泌体。通过体外实验,我们将利用RT-PCR、qPCR和免疫印迹技术分析前hascs对tgf - β信号的抑制作用。利用显微ct、组织学和免疫荧光染色在颅骨缺损模型中分析前hascs的骨骨再生能力。分离的外泌体的货物含量将通过成像技术、免疫印迹、蛋白质组学分析和miRNA分析来表征tgf - β信号与未经处理的hASCs外泌体的抑制。最后,我们将对候选蛋白和/或mirna进行功能测试,我们之前通过功能增益和功能丧失实验确定了这些候选蛋白和/或mirna。我们相信外泌体可以作为靶向药物递送的载体,并且可以通过基因预处理来改善靶部位的治疗。因此,外泌体有可能取代基于细胞的治疗,并消除MSC来源细胞移植的肿瘤转化风险。

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 }}

Dr. Julika Leandra Huber其他文献

Dr. Julika Leandra Huber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 }}

相似海外基金

Intranasal Delivery of Telomerase Reverse Transcriptase mRNA for Therapy ofTraumatic Brain Injury
鼻内递送端粒酶逆转录酶 mRNA 用于治疗创伤性脑损伤
  • 批准号:
    10602034
  • 财政年份:
    2022
  • 资助金额:
    --
  • 项目类别:
Study on the molecular basis that defines organ regenerative ability in an anuran amphibian (Xenopus laevis)
无尾两栖动物(非洲爪蟾)器官再生能力的分子基础研究
  • 批准号:
    20K21517
  • 财政年份:
    2020
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Challenging Research (Exploratory)
Elucidation of the tissue repair ability of CD163 positive macrophages aimed at regenerative medicine
阐明CD163阳性巨噬细胞的组织修复能力,旨在再生医学
  • 批准号:
    18J01441
  • 财政年份:
    2018
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for JSPS Fellows
Evoking regenerative ability based on the priciple of regeneration
基于再生原理激发再生能力
  • 批准号:
    16H06376
  • 财政年份:
    2016
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (S)
Development of regenerative medical technology for cartilage and meniscus using human synovial stem cells by reproducing natural healing ability of rodents
通过复制啮齿类动物的自然愈合能力,利用人类滑膜干细胞开发软骨和半月板再生医疗技术
  • 批准号:
    16H06262
  • 财政年份:
    2016
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Young Scientists (A)
Search for new specific molecular markers that can predict bone regenerative ability of mandible bone marrow stem cells.
寻找新的特异性分子标记来预测下颌骨骨髓干细胞的骨再生能力。
  • 批准号:
    16K11605
  • 财政年份:
    2016
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Development of regenerative medical materials with the ability to promote wound healing using CXCL12.
使用 CXCL12 开发能够促进伤口愈合的再生医疗材料。
  • 批准号:
    16K11635
  • 财政年份:
    2016
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Evolutionary loss of regenerative ability: The role of evolutionary conserved regeneration signal-response enhancer
再生能力的进化丧失:进化保守的再生信号反应增强子的作用
  • 批准号:
    16K07362
  • 财政年份:
    2016
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Study on the reactivation of regenerative ability by masking 'autoantigens' expressed in organ blastema.
通过掩蔽器官胚基中表达的“自身抗原”来重新激活再生能力的研究。
  • 批准号:
    26640051
  • 财政年份:
    2014
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Attempts to acquire the regenerative ability by use of epigenetic regulation in the damaged mouse retina
尝试通过表观遗传调控受损小鼠视网膜获得再生能力
  • 批准号:
    26462699
  • 财政年份:
    2014
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了