EXOSOMES: ROLE IN ALLOGRAFT REJECTION AND POTENTIAL AS A BIOMARKER

外泌体：在同种异体移植排斥中的作用以及作为生物标志物的潜力

• 批准号: 9243980
• 负责人: THALACHALLOUR MOHANAKUMAR
• 金额: $ 18.59万
• 依托单位: ST. JOSEPH'S HOSPITAL AND MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243980
• 关键词: Acute; Antibodies; Area; Binding; Biological Markers; Biopsy; Biopsy Specimen; Blood Circulation; Body Fluids; Bronchiolitis; Bronchiolitis Obliterans; Bronchoalveolar Lavage Fluid; Cells; Characteristics; Chronic; Chronic Obstructive Airway Disease; Classification; Clinical; Collagen; Consensus; Cystic Fibrosis; Data; Detection; Development; Diagnosis; Diagnostic; Disease; Endothelial Cells; Epithelial; Epithelial Cells; Evaluation; Event; Extracellular Matrix Proteins; Fibrosis; Gene Expression; Goals; Gold; Hamman-Rich syndrome; Histologic; Human; Immune Targeting; Immune response; Impairment; Interobserver Variability; Kinetics; Lead; Lesion; Leucocytic infiltrate; Lung; Lung Transplantation; Lung diseases; Lymphocyte; Measurement; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular Profiling; Organ; Organ Transplantation; Pathology; Phenotype; Proteins; Reader; Resistance; Role; Serum; Site; Spirometry; Structure of parenchyma of lung; Structure of respiratory bronchiole; Syndrome; Terminal Bronchiole; Testing; Transplant Recipients; Ultracentrifugation; allograft rejection; base; diagnosis standard; exosome; experimental study; immunoregulation; improved; insight; lung allograft; mouse model; novel; potential biomarker; public health relevance; selective expression; specific biomarkers

 DESCRIPTION (provided by applicant): Lung transplantation (LTx) is a viable treatment option for end-stage lung diseases. Although short-term survival has improved, acute and chronic rejection remains as hurdles for long-term function of the organ. Diagnosis of both acute and chronic rejection following human LTx remains an important challenge. Transbronchial lung biopsy has been the gold standard for the diagnosis of acute rejection. However, there is significant interobserver variability, and up to 40% of biopsies are insufficient for evaluation of rejection. Diagnostic features for Ab mediated rejection also varies widely. Lastly, chronic rejection after LTx is characterized histologically by obliterative bronchiolitis (OB), a fibroproliferative lesion involving terminal and respiratory bronchioles. Transbronchial lung biopsy is an insensitive method for the detection of OB. Therefore, chronic lung allograft rejection is diagnosed and staged according to decrements in spirometry measurements, a downstream event that induces airway fibrosis and obliteration. Therefore, further insights into the mechanisms of rejection and identifying specific biomarkers for rejection after LTx is critical for long term function of the transplanted lungs. Using both human LTxR and a murine model of chronic rejection, obliterataive airway disease (OAD), we demonstrated that: 1) In human LTxR, circulating exosomes are detectable during acute cellular rejection (A1 and A2). 2) Exosomes can be detected in the sera earlier to acute cellular rejection. 3) De novo development of DSA is accompanied by exosomes in the bronchoalveolar lavage fluid (BAL). 4) Exosomes contained Collagen V (Col-V) which has been shown to be selectively expressed in the lung parenchyma. 5) Exosomes are present in the local site (BAL) much before clinical evidences of OAD lesions, and 6) Sera from LTxR diagnosed with BOS also have Col-V containing exosomes which are detectable in the sera much before BOS. Based on these findings, we propose to determine; 1) The mechanisms by which exosome induction following donor specific immune responses lead to lung allograft rejection; and 2) Determine the kinetics of exosome development with the goal to employ exosome detection in the sera as a potential biomarker for rejection and treatment of rejection. Towards this, we will determine the kinetics of exosomes present in the BAL and serum following LTx using ultracentrifugation and protein isolation methods and correlate the findings with acute cellular, acute Ab mediated, and chronic rejection. Subsequent experiments will; a) define the composition of exosomes; b) determine the cells contributing to the exosome; and c) determine whether HLA specific messenger RNA (mRNA) present in the exosomes will transfer the donor HLA to recipient epithelial and endothelial cells. This is based on our premise that the exosomes will contain several microRNAs involved in immune regulation and donor HLA mRNA. Further, mRNA for HLA in the exosomes may transfer the message to the recipient's epithelial or endothelial cells leading to continued immune responses against the donor HLA resulting in chronic rejection following human LTx.

 描述（由申请人提供）：肺移植（LTx）是终末期肺病的可行治疗选择，尽管短期生存有所改善，但急性和慢性排斥仍然是器官长期功能诊断的障碍。人类 LTx 后的急性和慢性排斥反应仍然是一个重要的挑战。经支气管肺活检一直是诊断急性排斥反应的金标准，然而，观察者之间存在显着差异，高达 40% 的活检结果存在显着差异。不足以评估 Ab 介导的排斥反应的诊断特征也很广泛。 ，慢性肺同种异体移植排斥反应是根据肺活量测量的减少来诊断和分期的，这是一种诱导气道纤维化的下游事件，因此，进一步了解 LTx 后的排斥机制并确定排斥的特定生物标志物至关重要。 对于移植肺的长期功能，我们使用人类 LTxR 和慢性排斥、闭塞性气道疾病 (OAD) 的小鼠模型证明：1) 在人类 LTxR 中，在急性细胞排斥期间可检测到循环外泌体（A1 和 A2）。 ). 2) 急性细胞排斥反应早期可在血清中检测到外泌体。 3) DSA 的从头发育伴随着支气管肺泡中的外泌体。 4) 外泌体含有 V 型胶原蛋白 (Col-V)，已被证明在肺实质中选择性表达。 5) 外泌体早在 OAD 病变的临床证据之前就存在于局部部位 (BAL)， 6) 诊断为 BOS 的 LTxR 血清也含有含有外泌体的 Col-V，这些外泌体在 BOS 之前就可以在血清中检测到。 1) 供体特异性免疫反应后外泌体诱导导致肺同种异体移植排斥的机制；2) 确定外泌体发育的动力学，目的是利用血清中的外泌体检测作为排斥反应和排斥反应治疗的潜在生物标志物。为此，我们将使用超速离心和蛋白质分离方法确定 LTx 后 BAL 和血清中存在的外泌体的动力学，并将结果与​​急性细胞、急性 Ab 介导的和慢性排斥反应相关联。 a) 确定外泌体的组成；b) 确定形成外泌体的细胞；以及 c) 确定外泌体中存在的 HLA 特异性信使 RNA (mRNA) 是否会将供体 HLA 转移至受体上皮细胞和内皮细胞。基于我们的假设，外泌体将含有几种参与免疫调节的 microRNA 和供体 HLA mRNA，此外，外泌体中的 HLA mRNA 可能会将信息传递给受体的上皮细胞或受体。内皮细胞导致针对供体 HLA 的持续免疫反应，导致人类 LTx 后的慢性排斥反应。

项目成果

Extracellular Vesicles Mediate Immune Responses to Tissue-Associated Self-Antigens: Role in Solid Organ Transplantations.

细胞外囊泡介导对组织相关自身抗原的免疫反应：在实体器官移植中的作用。

• 发表时间: 2022
• 作者: Ravichandran, Ranjithkumar;Bansal, Sandhya;Rahman, Mohammad;Sureshbabu, Angara;Sankpal, Narendra;Fleming, Timothy;Bharat, Ankit;Mohanakumar, Thalachallour
• 通讯作者: Mohanakumar, Thalachallour

Low-dose IL-2 prevents murine chronic cardiac allograft rejection: Role for IL-2-induced T regulatory cells and exosomes with PD-L1 and CD73.

低剂量 IL-2 可预防小鼠慢性心脏同种异体移植排斥：IL-2 诱导的 T 调节细胞和外泌体与 PD-L1 和 CD73 的作用。

• 发表时间: 2022-09
• 作者: Ravichandran, Ranjithkumar;Itabashi, Yoshihiro;Fleming, Timothy;Bansal, Sandhya;Bowen, Sara;Poulson, Christin;Bharat, Ankit;Bremner, Ross;Smith, Michael;Mohanakumar, Thalachallour
• 通讯作者: Mohanakumar, Thalachallour

Chronic Lung Allograft Dysfunction: Immune Responses Induced by Circulating Exosomes with Lung-Associated Self-Antigens.

慢性肺同种异体移植功能障碍：循环外泌体与肺相关自身抗原诱导的免疫反应。

• 发表时间: 2019
• 影响因子: 1.3
• 作者: Rahman, Mohammad;Sureshbabu, Angara;Tokman, Sofya;Mohanakumar, Thalachallour
• 通讯作者: Mohanakumar, Thalachallour

Role for exosomes with self-antigens and immune regulatory molecules in allo- and auto-immunity leading to chronic immune injury following murine kidney transplantation.

具有自身抗原和免疫调节分子的外泌体在同种免疫和自身免疫中的作用，导致小鼠肾移植后的慢性免疫损伤。

• DOI: 10.1016/j.trim.2022.101702
• 发表时间: 2022-08-01
• 期刊: Transplant immunology
• 影响因子: 1.5
• 作者: Y. Itabashi;R. Ravich;ran;ran;S. Bansal;Chiahsuan Chin;C. Poulson;A. Sureshbabu;S. Nair;S. Perincheri;T. Mohanakumar
• 通讯作者: T. Mohanakumar

A decline in club cell secretory proteins in lung transplantation is associated with release of natural killer cells exosomes leading to chronic rejection.

肺移植中俱乐部细胞分泌蛋白的下降与导致慢性排斥的自然杀伤细胞外泌体的释放有关。

• 发表时间: 2021-12
• 作者: Ravichandran, Ranjithkumar;Itabashi, Yoshihiro;Liu, Wei;Bansal, Sandhya;Rahman, Mohammad;Poulson, Christin;Fleming, Timothy;Bremner, Ross M;Smith, Michael;Mohanakumar, Thalachallour
• 通讯作者: Mohanakumar, Thalachallour