Biomolecular Markers for Safe Minimization of Immunosuppression

用于安全最小化免疫抑制的生物分子标记

• 批准号: 10209348
• 负责人: MANIKKAM SUTHANTHIRAN
• 金额: $ 37.49万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-28 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10209348
• 关键词: 2019-nCoV; Acute Renal Failure with Renal Papillary Necrosis; Address; Administrative Supplement; Admission activity; Allografting; Bioinformatics; Biological Assay; Biopsy; CD3E gene; COVID-19; COVID-19 diagnosis; COVID-19 outbreak; COVID-19 patient; CXCL10 gene; Cells; Clinical; Cohort Studies; Collection; Complementary DNA; Creatinine; Custom; Diagnosis; Diagnostic; Equation; Functional disorder; Funding; Gene Expression Profiling; Genes; Goals; Grant; Health Personnel; Hemorrhage; Home; Hospitalization; Hour; Immune; Immune response; Immunosuppression; Kidney Transplantation; Measurement; Measures; Messenger RNA; Methods; Molecular; Monitor; Outcome; Patient Recruitments; Patients; Procedures; RNA; RNA Sequences; Renal function; Research; Retrieval; Ribosomal RNA; Risk; SARS-CoV-2 positive; SARS-CoV-2 transmission; Sampling; Serum; Specimen; Symptoms; Therapeutic immunosuppression; Time; Training; Urine; Validation; base; biobank; cohort; genetic signature; innovation; insight; kidney allograft; prognostic; recruit; response; severe COVID-19; symptomatic COVID-19; symptomatology; tool; transcriptome; transcriptome sequencing; transcriptomics; urinary

We seek additional funds for research responsive to the SARS-CoV-2/COVID-19 outbreak that is in scope of our ongoing grant R37AI051652 “Biomolecular Markers for Safe Minimization of Immuno-suppression.” From March 13, 2020 to April 20, 2020, we hospitalized 39 kidney allograft recipients positive for SARS-CoV-2 and with Covid-19 symptoms. Among these patients, 20 (51%) developed acute kidney injury (AKI). Importantly, graft dysfunction due to AKI recovered in 9 patients only and did not recover in 11 patients as of May 15, 2020. None underwent a diagnostic allograft biopsy because of biopsy-associated complications such as bleeding are potentially much more serious in this cohort and also to limit potential exposure of healthcare workers to SARS- CoV-2 during the invasive biopsy procedure. In the absence of a diagnostic biopsy, none received anti-rejection therapy. Whether the graft dysfunction was reversible or nonreversible could not be predicted at the time of graft dysfunction diagnosis. The dynamics of anti-allograft response from the reductions in their immunosuppressive therapy could not be captured with the available clinical analytes. To address these existing challenges, we propose the following: Specific Aim 1. To perform RNA sequencing of urinary cells and investigate whether the urinary cell transcriptome, ascertained at the time of graft dysfunction, is prognostic of allograft dysfunction. Urine will be collected at the time of graft dysfunction diagnosis and RNA isolated from urinary cells. RNA from 30 patients with reversible graft dysfunction; RNA from 30 patients with nonreversible graft dysfunction; and RNA from 30 patients with no graft dysfunction during the 3 months since Covid-19 diagnosis will be RNA sequenced and bioinformatics performed. The goal is to determine whether the urinary cell transcriptome profile, ascertained at the time of graft dysfunction, is prognostic of graft dysfunction and distinguishes those with reversible graft dysfunction from those with nonreversible graft dysfunction. Specific Aim 2. To measure urinary cell 3-gene signature score in sequential urine samples from Covid-19 kidney allograft recipients. Urine will be collected at baseline and sequentially every two weeks for 3 months since Covid-19 diagnosis. We will retrieve 210 sequential samples from 30 Covid-19 kidney allograft recipients (Baseline and 6 sequential samples from each patient) who developed reversible graft dysfunction; 210 sequential samples from 30 Covid-19 kidney allograft recipients who developed nonreversible graft dysfunction; and 30 Covid-19 kidney allograft recipients who did not develop graft dysfunction during the 3-months since Covid-19 diagnosis. RNA isolated from urinary cells will be reverse transcribed to cDNA and absolute copy numbers of CD3E mRNA, CXCL10 mRNA and 18S rRNA will be measured using customized PCR assays and urinary cell 3-gene signature score will be computed using a validated regression equation. The objective is to determine whether the urinary cell 3-gene scores in sequential samples anticipate those who develop nonreversible graft dysfunction rom those who develop reversible graft dysfunction.

我们寻求额外的资金来响应SARS-COV-2/COVID-19爆发的研究，该爆发的范围 我们正在进行的赠款R37AI051652“安全最大程度地减少免疫抑制的生物分子标记”。从 2020年3月13日至2020年4月20日，我们住院的39名肾脏同种异体接受者对SARS-COV-2和 有199症状。在这些患者中，有20名（51％）出现急性肾脏损伤（AKI）。重要的是，移植物 由于AKI的功能障碍仅在9例患者中恢复，但截至2020年5月15日，11例患者没有康复。 由于活检相关并发症（例如出血），接受了同种异体移植活检 在这个队列中，可能会更加严重，也可以限制医护人员的潜在暴露于SARS- 在侵入性活检过程中的COV-2。在没有诊断活检的情况下，没有人接受反拒绝 治疗。在移植时无法预测移植功能障碍是可逆的还是不可逆的 功能障碍诊断。免疫抑制作用减少的抗同盟移植反应的动力学 无法使用可用的临床分析物来捕获治疗。为了应对这些现有挑战，我们 提案以下：特定目的1。进行尿细胞的RNA测序并研究 在移植功能障碍时确定的尿细胞转录组是否为预后 同种异体功能障碍。将在移植功能障碍诊断时收集尿液，并从中分离出RNA 泌尿细胞。 30例可逆移植功能障碍患者的RNA；来自30例不可逆患者的RNA 移植功能障碍；在Covid-19以来的3个月中，30例没有移植功能障碍的患者的RNA和RNA 诊断将是RNA测序并进行生物信息学。目的是确定尿液是否 在移植功能障碍时确定的细胞转录组谱是移植功能障碍的预后和 将具有可逆移植功能障碍的人与具有非可逆移植功能障碍的人区分开。具体的 目的2。测量来自Covid-19肾脏顺序尿液样品中的尿细胞3基因签名评分 同种异体移植者。尿液将在基线时，并依次每两周收集一次，持续3个月 COVID-19诊断。我们将从30 Covid-19肾脏同种异体移植者中检索210个顺序样品 （来自每个患者的基线和6个顺序样品），患有可逆的移植功能障碍； 210 来自30个Covid-19-19肾脏同种异体受体的顺序样品，患有非可逆移植功能障碍； 和30名Covid-19肾脏同种异体接受者，他们在3个月以来没有出现移植功能障碍 COVID-19诊断。从泌尿细胞中分离的RNA将被反转录为cDNA和绝对拷贝 CD3E mRNA，CXCL10 mRNA和18S rRNA的数量将使用自定义的PCR分析和 将使用经过验证的回归方程计算尿中3-GENE签名评分。目标是 为了确定顺序样品中的尿中3基分数是否预期 不可逆的移植功能障碍ROM那些患有可逆移植功能障碍的人。

项目成果

Urinary cell-free DNA is a versatile analyte for monitoring infections of the urinary tract.

• DOI: 10.1038/s41467-018-04745-0
• 发表时间: 2018-06-20
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Burnham P;Dadhania D;Heyang M;Chen F;Westblade LF;Suthanthiran M;Lee JR;De Vlaminck I
• 通讯作者: De Vlaminck I

Gut microbiota and tacrolimus dosing in kidney transplantation.

• DOI: 10.1371/journal.pone.0122399
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Lee JR;Muthukumar T;Dadhania D;Taur Y;Jenq RR;Toussaint NC;Ling L;Pamer E;Suthanthiran M
• 通讯作者: Suthanthiran M

Sex and Kidney Transplantation: Why Can't a Woman Be More Like a Man?

性与肾移植：为什么女人不能更像男人？

• DOI: 10.1681/asn.2017060657
• 发表时间: 2017
• 期刊: Journal of the American Society of Nephrology : JASN
• 作者: August,Phyllis;Suthanthiran,Manikkam
• 通讯作者: Suthanthiran,Manikkam

Detection of infiltrating fibroblasts by single-cell transcriptomics in human kidney allografts.

• DOI: 10.1371/journal.pone.0267704
• 发表时间: 2022
• 期刊: PLOS ONE
• 影响因子: 3.7
• 作者: Suryawanshi, Hemant;Yang, Hua;Lubetzky, Michelle;Morozov, Pavel;Lagman, Mila;Thareja, Gaurav;Alonso, Alicia;Li, Carol;Snopkowski, Catherine;Belkadi, Aziz;Mueller, Franco B.;Lee, John R.;Dadhania, Darshana M.;Salvatore, Steven P.;Seshan, Surya, V;Sharma, Vijay K.;Suhre, Karsten;Suthanthiran, Manikkam;Tuschl, Thomas;Muthukumar, Thangamani
• 通讯作者: Muthukumar, Thangamani

Gut microbiota dysbiosis and diarrhea in kidney transplant recipients.

• DOI: 10.1111/ajt.14974
• 发表时间: 2019-03
• 期刊: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons
• 作者: Lee JR;Magruder M;Zhang L;Westblade LF;Satlin MJ;Robertson A;Edusei E;Crawford C;Ling L;Taur Y;Schluter J;Lubetzky M;Dadhania D;Pamer E;Suthanthiran M
• 通讯作者: Suthanthiran M