21EBTA Driving Pluripotent Stem Cell Osteogenesis with Light for Tissue Engineering

21EBTA 利用光驱动组织工程多能干细胞成骨

• 批准号: BB/W013940/1
• 负责人: Susan Kimber
• 金额: $ 45.9万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW013940%2F1
• 关键词: 21EBTA; Driving; Pluripotent; Stem; Cell

Skeletal problems affecting bones and joints, such as fractures that don't heal or osteoarthritis are a major cause of disfunction, pain and disability in the over 45s. Human pluripotent stem cell can make different tissues, including bone, if the right reagents are added to the cells in a dish at appropriate times. However, generating such tissues is costly, relying on addition of proteins called growth factors- in particular one called bone morphogenetic protein (BMP). BMPs are important in development for making bone, but the protein is expensive and different batches have different activity, reducing the reproducibility of protocols. We will replace the use of BMP in our protocol by engineering stem cells to contain receptors which respond to a particular wavelength of visible light, instead of the BMP protein, to trigger the effect of BMP on the cells. We will evaluate the cell-response to light, in terms of intensity and timing of light pulses, to see if it replicates what the BMP protein does to the cells. As well as saving costs, this will give us much more accurate control of the bone-encouraging signals, compared to that obtainable by daily addition of the protein which is broken down by the cells. We will thus develop a novel method, driving the stem cells to form bone using light. First this will be done in a dish but then we will transfer this to a 3 dimensional format; we will use a 3D printer to print the light responsive cells to make a 3D construct by incorporating them into a gel containing collagen, found in bone, and hydroxyapatite which will encourage mineralisation of the bone. We will combine this with a stiffer scaffold to encourage bone formation. We will monitor the printing and culture parameters to give the most authentic bone tissue and characterise this. Having established this system, in the future we will take this forward in 3 ways 1) to make bone-like constructs suitable for healing bone lesions; 2) to combine with light driven, pluripotent stem cell-generated, cartilage cells and make a construct which can be used to investigate the formation of the joint and factors causing joint disease; 3) to use as a model system for discovering new drugs which enhance bone healing or correct bone or joint disease abnormalities. These light-driven engineered cells can also be applied to other BMP-dependent human stem cell generated tissues, for understanding development and disease, and the pipeline can be used to engineer other growth factor responses applicable to human health and manufacturing processes. Thus this marks some of the first step in this promising area of synthetic biology, opening up the use of light with engineered human pluripotent stem cells and bioprinting to drive different applications.

影响骨骼和关节的骨骼问题，如骨折无法愈合或骨关节炎，是45岁以上患者功能障碍、疼痛和残疾的主要原因。如果在适当的时间将合适的试剂加入培养皿中的细胞中，人类多能干细胞可以形成包括骨骼在内的不同组织。然而，生成这样的组织是昂贵的，依赖于添加称为生长因子的蛋白质--特别是一种名为骨形态发生蛋白(BMP)的蛋白质。BMP在骨的开发中很重要，但蛋白质价格昂贵，不同批次的活性不同，降低了方案的重复性。我们将取代在我们的方案中使用BMP，通过改造干细胞含有对特定波长的可见光做出反应的受体，而不是BMP蛋白，以触发BMP对细胞的影响。我们将根据光脉冲的强度和时间来评估细胞对光的反应，看看它是否复制了BMP蛋白对细胞的作用。除了节省成本，与每天添加被细胞分解的蛋白质相比，这将使我们能够更准确地控制骨骼激励信号。因此，我们将开发一种新的方法，利用光驱动干细胞形成骨。首先，这将在培养皿中完成，然后我们将把它转换成3D格式；我们将使用3D打印机打印光响应细胞，通过将它们结合到含有胶原蛋白的凝胶中来构建3D结构，胶原蛋白存在于骨骼中，羟基磷灰石将促进骨骼的矿化。我们将把它与更坚硬的支架结合起来，以鼓励骨形成。我们将监测印刷和培养参数，以获得最真实的骨组织并对其进行表征。在建立了这个系统之后，未来我们将从3个方面进行推进：1)制造适合于修复骨骼损伤的骨样结构；2)与光驱动的多能干细胞生成的软骨细胞相结合，制造可用于研究关节形成和导致关节疾病的因素的结构；3)作为发现促进骨愈合或纠正骨或关节疾病异常的新药的模型系统。这些光驱动的工程细胞也可以应用于其他依赖BMP的人类干细胞产生的组织，以了解发育和疾病，该管道可以用于设计其他适用于人类健康和制造过程的生长因子反应。因此，这标志着合成生物学这一前景看好的领域迈出了一些第一步，开启了光与工程人类多能干细胞的使用，并通过生物打印来驱动不同的应用。

项目成果

Development of human cartilage circadian rhythm in a stem cell-chondrogenesis model.

• DOI: 10.7150/thno.70893
• 发表时间: 2022
• 期刊: THERANOSTICS
• 影响因子: 12.4
• 作者: Naven, Mark A.;Zeef, Leo A. H.;Li, Shiyang;Humphreys, Paul A.;Smith, Christopher A.;Pathiranage, Dharshika;Cain, Stuart;Woods, Steven;Bates, Nicola;Au, Manting;Wen, Chunyi;Kimber, Susan J.;Meng, Qing-Jun
• 通讯作者: Meng, Qing-Jun

Optogenetic manipulation of BMP signaling to drive chondrogenic differentiation of hPSCs

• DOI: 10.1016/j.celrep.2023.113502
• 发表时间: 2023-11-28
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Humphreys，Paul E. A.;Woods，Steven;Kimber，Susan J.
• 通讯作者: Kimber，Susan J.

SIRT1 activity orchestrates ECM expression during hESC-chondrogenic differentiation.

• DOI: 10.1096/fj.202200169r
• 发表时间: 2022-05
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Smith CA;Humphreys PA;Bates N;Naven MA;Cain SA;Dvir-Ginzberg M;Kimber SJ
• 通讯作者: Kimber SJ

Effect of a retinoic acid analogue on BMP-driven pluripotent stem cell chondrogenesis

• DOI: 10.1101/2023.06.20.545738
• 发表时间: 2023-06
• 期刊: bioRxiv
• 作者: F. Mancini;Paul A Humphreys;Steven Woods;N. Bates;Sara Cuvertino;Julieta O’Flaherty;Leela C. Biant;M. Domingos;S. Kimber

Effect of a retinoic acid analogue on BMP-driven pluripotent stem cell chondrogenesis.

• DOI: 10.1038/s41598-024-52362-3
• 发表时间: 2024-02-01
• 期刊: Scientific reports
• 影响因子: 4.6