Human Specimen Collection to Support Basic and Clinical Research

支持基础和临床研究的人体样本采集

基本信息

项目摘要

In Dr Shets and Dr. Theins labs, blood samples were obtained from healthy volunteers and sickle cell patients to further studies on: 1. Platelet Activation in Sickle Cell Disease: Platelets were isolated from whole blood, to study in vitro platelet aggregation studies. Whole blood was used to study platelet aggregation and ATP luminescence. 2. Tissue Damage and Release of Cell-Free DNA: Optimization of cell-free DNA extraction, quantitation and library preparation methodology. Whole genome sequencing (WGS) and whole genome bisulfite sequencing (WGBS), and quantitative polymerase chain reaction (qPCR), where the cell-free DNA extracted from healthy volunteers was used as an independent healthy control to compare with sickle cell disease patients who are in steady-state and crisis. Absence of mitochonidrial DNA in RBCs was confirmed in healthy volunteers. 3. Neutrophil Activation and Extracellular Trap Formation: Blood from healthy volunteers and plasma from sickle cell patients were used for in vitro neutrophil functional studies (neutrophils extracellular traps formation). In separate experiments blood from both healthy donors and sickle cells patients were used to isolate low density granulocytes (LDGs) using density gradient centrifugation and flow cytometry. 4. Extracellular vesicles (EVs): Isolation of EVs from plasma obtained from anti coagulated blood samples of SCD patients and Healthy volunteers for microparticle. These assays are being standardized for two newly established protocols to study venous thromboembolism and an antithrombotic agent, Isoquercetin. 5. Platelet Activation: Blood from healthy volunteers was used for in vitro platelet activation studies (thrombin generation and blood coagulation). In Dr. Tisdales lab, studies focused on developing genetic strategies aimed at correction of SCD through modification of autologous hematopoietic stem cells (HSCs) from the marrow. These furthered optimization of HSCs for gene therapy. 1. Collection of bone marrow HSCs is being optimized in an ongoing clinical trial testing lentiviral gene transfer to HSCs in patients with SCD. Therefore, bone marrow continues to be collected from volunteer patients to optimize cell processing and HSC enrichment. Twenty milliliters of BM from subjects with SCD (HbSS genotype) was collected in different anticoagulants (Heparin, ACD-A) and processed immediately (day 0) or stored at 4 degrees C and processed the following day (day 1). After isolation via Ficoll density gradient centrifugation, the mononuclear (MN) layer was stained with antibodies against inflammatory markers (CD36, CD35, CD11b, CD62L, CD62P), non-MN cells (GPA, CD66b, CD41/61), or processed for CD34+ selection using a magnetic microbead CD34+ selection kit and stained for CD34, CD45, and GPA expression. 2. Data were analyzed by conventional and imaging flow cytometry, the latter confirming post-CD34+ selection flow data and demonstrating antibody intensity as a characterization of HSC heterogeneity and progenitor lineage. In Dr Eatons lab, blood has been obtained from sickle cell patients to develop assays that quantify the rate of sickling and the effects of cell volume, hemoglobin concentration on sickling kinetics. 1. These approaches are being used to test the effects of anti-sickling drugs and to study the effects of human variation in iron availability, on the rate of sickling. Further studies are ongoing in this regard. Specifically, the effect of anti-sickling drugs are being evaluated in healthy controls and those with sickle cell trait. 2. Studies to standardize assays to measure oxygen equilibrium curves in the presence of antisickling agents are also underway. In Dr Fitzhughs lab, blood has been obtained from sickle cell patients and healthy controls to standardize cytokine measurements in SCD patients who underwent haploidentical hematopoietic stem cell transplantation. In particular, these measurements included assays for thrombospondin and platelet factor 4 which helped interpret plasma and serum biomarkers of successful and failed engraftment.
在谢茨博士和泰恩斯博士的实验室里,他们从健康志愿者和镰状细胞病患者身上采集了血液样本,以进一步研究:

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)
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Arun Shet其他文献

Arun Shet的其他文献

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{{ truncateString('Arun Shet', 18)}}的其他基金

Human Specimen Collection to Support Basic and Clinical Research
支持基础和临床研究的人体样本采集
  • 批准号:
    10492971
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
Venous Thrombosis Biomarkers in Sickle Cell Disease and Sickle Cell Trait
镰状细胞病和镰状细胞性状中的静脉血栓形成生物标志物
  • 批准号:
    10262685
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
A Study to Evaluate the Effects of fixed dose Flavonoid Isoquercetin on thrombo-inflammatory biomarkers in subjects with stable Sickle Cell Disease
评估固定剂量类黄酮异槲皮素对稳定镰状细胞病患者血栓炎症生物标志物影响的研究
  • 批准号:
    10492974
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
A Study to Evaluate the Effects of fixed dose Flavonoid Isoquercetin on thrombo-inflammatory biomarkers in subjects with stable Sickle Cell Disease
评估固定剂量类黄酮异槲皮素对稳定镰状细胞病患者血栓炎症生物标志物影响的研究
  • 批准号:
    10929196
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
Venous Thrombosis Biomarkers in Sickle Cell Disease and Sickle Cell Trait
镰状细胞病和镰状细胞性状中的静脉血栓形成生物标志物
  • 批准号:
    10492973
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
Venous Thrombosis Biomarkers in Sickle Cell Disease and Sickle Cell Trait
镰状细胞病和镰状细胞性状中的静脉血栓形成生物标志物
  • 批准号:
    10706189
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
A Study to Evaluate the Effects of fixed dose Flavonoid Isoquercetin on thrombo-inflammatory biomarkers in subjects with stable Sickle Cell Disease
评估固定剂量类黄酮异槲皮素对稳定镰状细胞病患者血栓炎症生物标志物影响的研究
  • 批准号:
    10706190
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
Human Specimen Collection to Support Basic and Clinical Research
支持基础和临床研究的人体样本采集
  • 批准号:
    10706185
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
Human Specimen Collection to Support Basic and Clinical Research
支持基础和临床研究的人体样本采集
  • 批准号:
    10929189
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:
A Phase 1 Study to Evaluate the Safety and Tolerability of Escalating Doses of Fostamatinib in Subjects with stable sickle cell disease
评估 Fostamatinib 剂量递增对稳定镰状细胞病受试者的安全性和耐受性的 1 期研究
  • 批准号:
    10930552
  • 财政年份:
  • 资助金额:
    $ 31.68万
  • 项目类别:

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