An ex Vivo Study to Purify Amyloid Fibrils and Solve Their 3D Structures Using Amyloid Proteins From Medullary Thyroid Cancer and Laryngeal Amyloidosis
试验速览
- 阶段
- 不适用
- 状态
- 尚未招募
- 发起方
- 入组人数
- 10
- 主要终点
- Transmission electron microscopy
研究概览
简要总结
Using excess tumour samples that contain amyoid, from patients with Medullary Thyroid Cancer, we aim to determine the structures of ex vivo amyloid fibrils from human tumour tissue samples and compare them with that of existing stock of in vitro formed amyloid fibrils. This will permit the analysis of the effects of gene mutation and post-translational modification on the development of amyloid from a disease state.
Amyloid is known to accumulate in the brain tissue of patients with neuro-degenerative conditions such as Alzheimer's disease and Dementia. Therefore solving the structure of amyloid fibrils may aid the development of future treatments for these conditions.
详细描述
This is an anonymous, voluntary inclusion, laboratory based, basic science research study. Appropriate patients who meet the inclusion criteria and complete the consent process will be asked to donate a small sample of excess tumour tissue after the completion of their surgical procedure. This tissue will be transferred to the research laboratory in Leeds University for analysis and subject to a variety of investigations and scientific procedures to achieve the study objectives.
These include:
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- amyloid extraction by tissue homogenization and centrifugation as already documented in the literature.
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- mass spectrometry analysis to identify and characterize the most prominent proteins present in the amyloid deposits and their possible post-translational modifications.
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- Transmission Electron Microscopy to characterize at the gross features of the extracted amyloid fibrils at low resolution (>20Å). This relatively simple procedure allows for quick characterization of the samples in order to optimize them for Atomic Force Microscopy (AFM) and Cryo-Electron Microscopy (Cryo-EM).
-
- Atomic Force microscopy to characterize helical parameters of the amyloid fibrils needed to calculate the structure from Cryo-EM data.
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- Cryo-Electron Microscopy to elucidate the structure of the amyloids extracted from tissue affected in each disease
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Diagnosed with known or suspected MTC or LA
- •Age >= 18 years, no upper age limit
- •Able to provide informed consent for both surgical treatment and inclusion into this study
- •For MTC patients, treatment recommendation from the Thyroid MDT is for either hemithyroidectomy or total thyroidectomy
- •for LA patients, agreement with their attending clinician to undergo surgical debulking of disease under general anaesthetic. The procedure is called 'microlaryngoscopy and biopsy/debulking'
排除标准
- •No known or suspected diagnosis of MTC or LA
- •Age <18 years
- •Unable to provide informed consent
- •Treatment recommendation from Thyroid MDT is for any treatment excluding primary surgery (ie palliation, best supportive care, chemotherapy, radiotherapy etc)
- •For LA patients, no agreement between patient and attending clinician to proceed with surgery as primary treatment.
结局指标
主要结局
Transmission electron microscopy
时间窗: 12-24 months
To characterize the gross features of the extracted amyloid fibrils at low resolution (\>20Å). This relatively simple procedure allows for quick characterization of the samples in order to optimize them for Atomic Force Microscopy (AFM) and Cryo-Electron Microscopy (Cryo-EM).
Atomic force microscopy
时间窗: 12-24 months
To characterize helical parameters of the amyloid fibrils needed to calculate the structure from Cryo-EM data.
Cryo-electron microscopy (Cryo-EM)
时间窗: 12-24 months
To elucidate the structure of the amyloids extracted from tissue affected in each disease
Amyloid extraction by tissue homogenization and centrifugation
时间窗: 12-24 months
A standard laboratory technique for subsequent analysis
Mass spectrometry
时间窗: 12-24 months
Analysis to identify and characterize the most prominent proteins present in the amyloid deposits and their possible post-translational modifications.
次要结局
未报告次要终点