本文選題：分子影像學(xué) + 放射性核素光學(xué)成像��； 參考：《中國(guó)協(xié)和醫(yī)科大學(xué)》2010年博士論文

【摘要】： 分子影像學(xué)是一門新興的交叉學(xué)科,涉及到影像學(xué)、分子材料學(xué)(包括納米材料)、分子生物學(xué)(包括信號(hào)傳導(dǎo)通路、受體、抗體、配體等)、基因研究等。目前已有多種影像學(xué)技術(shù)應(yīng)用于分子影像學(xué)研究,如放射性核素成像(nuclear imaging),光學(xué)成像(optical imaging,OI),超聲成像(ultrosonic imaging)和核磁共振成像(Magnetic resonance imaging, MRI)等。放射性核素成像手段主要包括正電子發(fā)射計(jì)算機(jī)斷層成像(position emission tomography, PET)和單光子發(fā)射計(jì)算機(jī)斷層成像(single photon emission computed tomography, SPECT)。放射性核素成像首先將放射性藥物引入患者體內(nèi),在體外檢測(cè)放射性藥物發(fā)射出的高能量高穿透性的伽瑪射線,從而研究藥物在體內(nèi)的分布。通過(guò)與高分辨率的計(jì)算機(jī)斷層掃描技術(shù)(X-ray computed tomography)相結(jié)合,PET或SPECT可以在顯示深部組織的分子影像學(xué)特征的同時(shí),高分辨率地顯示組織的解剖結(jié)構(gòu),目前已廣泛地應(yīng)用于臨床。光學(xué)成像主要包括生物發(fā)光成像(bioluminescent imaging)、熒光成像(fluorescence imaging)等,應(yīng)用于分子及細(xì)胞生物學(xué)研究和活體(in vivo)表面成像。由于目前通過(guò)美國(guó)食品藥品管理局(Food and Drug Admistraton, FDA)認(rèn)證的光學(xué)探針只有吲哚菁綠(indocyanine green, ICG),光學(xué)成像在臨床應(yīng)用較少。多數(shù)活體光學(xué)成像只初步用于小動(dòng)物的實(shí)驗(yàn)成像。光學(xué)成像相比放射性核素成像價(jià)格較低廉,且允許具有不同光譜特征的探針進(jìn)行多通道成像。 本文在國(guó)際上率先綜合了核醫(yī)學(xué)示蹤劑和光學(xué)成像系統(tǒng)。使用光學(xué)系統(tǒng)的高敏感CCD相機(jī)檢測(cè)放射性探針以韌致輻射(Bremsstrahlung radiation)或契倫科夫輻射(Cerenkov radiation)產(chǎn)生的低能量光子,證明了核醫(yī)學(xué)探針應(yīng)用于光學(xué)成像的可行性,拓展了光學(xué)成像在臨床的應(yīng)用前景,并為直接監(jiān)測(cè)釔-90 (yttrium-90,90Y)等目前難以監(jiān)測(cè)的放射性治療用核素提供了有效手段。 一般而言,多聚肽比單體擁有更好的受體親和力和活性。本研究以多聚黑色素細(xì)胞刺激激素(a-Melanocyte-Stimulating Hormone,簡(jiǎn)稱a-MSH)類似肽和黑色素瘤為模型,利用正電子發(fā)射計(jì)算機(jī)斷層掃描技術(shù)(Positron emission tomography,簡(jiǎn)稱PET),對(duì)多聚肽的受體親和力和體內(nèi)腫瘤靶向能力進(jìn)行了系統(tǒng)的比較研究。 用多肽固相合成法合成了MSH類似肽單體(MSH1),二聚體(MSH2)和四聚體(MSH4),并分別與金屬螯合劑DOTA聯(lián)接(DOTA-MSH1, DOTA-MSH2, DOTA-MSH4)。通過(guò)B16/F10鼠黑色素瘤細(xì)胞測(cè)定三種多肽及聯(lián)接DOTA后的半數(shù)抑制濃度(IC50),評(píng)價(jià)其受體親和力。標(biāo)記64Cu后,進(jìn)行PET掃描成像,觀察其體內(nèi)分布和腫瘤顯像效果。并在不同時(shí)間點(diǎn)處死小鼠,取血、腫瘤及各主要器官,測(cè)量其每克組織注射百分劑量率(%ID/g)。多肽的各步反應(yīng)均使用反相高效液相色譜法(RP-HPLC)進(jìn)行純化并利用電噴霧質(zhì)譜(ESI-MS)進(jìn)行鑒定。DOTA-MSH4在體外實(shí)驗(yàn)中具有最高的受體親和力[IC50=1.00nM(95%可信區(qū)間0.83-1.21nM)],但在體內(nèi)實(shí)驗(yàn)中腫瘤攝取量最低(1小時(shí)=0.6l±0.02%ID/g；2小時(shí)=1.82±0.85%ID/g；4小時(shí)=2.98±0.24%ID/g),且在腎臟大量蓄積(1小時(shí)=12.91±1.86%ID/g；2小時(shí)=20.89±3.98%ID/g；4小時(shí)=24.98±2.17%ID/g)。相比之下,DOTA-MSH2有著中等的受體親和力[IC50=2.06nM(95%可信區(qū)間1.48-2.88nM)],卻有更高的腫瘤攝取量(1小時(shí)=5.65±1.13%ID/g；2小時(shí)=5.23±0.5%ID/g 0；4小時(shí)=5.39±0.84%ID/g),且在腎臟中的蓄積少于DOTA-MSH4(1小時(shí)=18.23±2.50%ID/g；2小時(shí)=16.84±3.57%ID/g；4小時(shí)=16.44±1.66%ID/g). DOTA-MSH1受體親和力最低[IC50=3.10nM(95%可信區(qū)間2.34-4.10nM)],有著中等的腫瘤攝取量(1小時(shí)=2.81+1.49%ID/g；2小時(shí)=3.74±1.57%ID/g；4小時(shí)=4.20±0.69%ID/g)和較低的腎臟蓄積(1小時(shí)=11.79±4.54%ID/g；2小時(shí)=11.24±3.95%ID/g；4小時(shí)=9.90±1.25%ID/g)。最后,對(duì)DOTA-MSH2進(jìn)行的體內(nèi)競(jìng)爭(zhēng)抑制實(shí)驗(yàn)顯示其具有良好的腫瘤特異性。高受體親和力的DOTA-MSH4卻有著較差的顯像效果。在本實(shí)驗(yàn)測(cè)試的三種多肽探針中,DOTA-MSH2被證明是最佳的黑色素瘤PET顯像劑。
[Abstract]:Molecular imaging is a new interdisciplinary subject involving imaging, molecular materials (including nanomaterials), molecular biology (including signal transduction pathways, receptors, antibodies, ligands, etc.), and gene studies. A variety of imaging techniques have been applied to molecular imaging studies, such as radionuclide imaging (nuclear imaging), Guang Xuecheng Such as (optical imaging, OI), ultrasound imaging (ultrosonic imaging) and nuclear magnetic resonance imaging (Magnetic resonance imaging, MRI), etc.. The radionuclide imaging means mainly include positron emission computed tomography (position emission tomography) and single photon emission computed tomography. Ography, SPECT). Radionuclide imaging first introduces radiopharmaceuticals into the patient and detects high energy and high penetration gamma rays emitted by radiopharmaceuticals in vitro, so as to study the distribution of drugs in the body. By combining with high resolution computed tomography (X-ray computed tomography), PET or SPECT can be used. In order to display the molecular imaging features of the deep tissue and display the anatomical structure of the tissue with high resolution, it is now widely used in clinic. Optical imaging mainly includes bioluminescent imaging, fluorescence imaging, and so on. It is applied to molecular and cellular biology research and living body (in vivo). Surface imaging. Because the optical probes certified by the Food and Drug Admistraton (FDA) are currently only indocyanine green (indocyanine green, ICG), optical imaging is less clinically used. Most of the living optical imaging is primarily used for experimental imaging of small animals. Optical imaging is compared to radionuclide imaging prices. It is cheaper and allows multi-channel imaging with probes with different spectral characteristics.
In this paper, nuclear medicine tracer and optical imaging system are first integrated in the world. Using the high sensitive CCD camera of the optical system to detect the low energy photon produced by the radioactive probe (Bremsstrahlung radiation) or the Cherenkov radiation (Cerenkov radiation), the feasibility of the application of the nuclear medical probe to optical imaging is demonstrated. It has expanded the prospect of clinical application of optical imaging and provides an effective means for the direct monitoring of yttrium -90 (yttrium-90,90Y), which is currently difficult to monitor in radionuclides.
In general, polypeptides have better receptor affinity and activity than monomers. This study uses polypeptides (a-Melanocyte-Stimulating Hormone, a-MSH) similar peptides and melanoma as models, and uses positron emission computed tomography (Positron emission tomography, PET) for polypeptides. The receptor affinity and in vivo tumor targeting ability were systematically compared.
MSH similar peptide monomer (MSH1), two polymer (MSH2) and four polymer (MSH4) were synthesized by the solid-phase peptide synthesis method, and DOTA coupling (DOTA-MSH1, DOTA-MSH2, DOTA-MSH4) with metal chelating agent (DOTA-MSH1, DOTA-MSH2, DOTA-MSH4) respectively. Three kinds of polypeptide and half inhibitory concentration (IC50) after joining DOTA were measured by B16/F10 rat melanoma cells, and the receptor affinity was evaluated. After labeling 64Cu, PET scanning was carried out to observe the distribution of the body and the effect of tumor imaging. The mice were killed at different time points, blood, tumor and the main organs were taken to measure the dose rate (%ID/g) per gram of tissue injection. All the steps of the peptide were purified by Reversed Phase High Performance liquid chromatography (RP-HPLC) and used by electrospray ionization mass spectrometry (ESI-MS). The identification of.DOTA-MSH4 has the highest receptor affinity [IC50=1.00nM (95% confidence interval 0.83-1.21nM) in vitro, but in the body, the tumor uptake is the lowest (1 hours =0.6l + 0.02%ID/g; 2 hours =1.82 + 0.85%ID/g; 4 hours =2.98 + 0.24%ID/g), and in a large amount of kidney accumulation (1 hours =12.91 + 1.86%ID/g; 2 hours =20.89) D/g; 4 hours =24.98 + 2.17%ID/g). In contrast, DOTA-MSH2 has a moderate receptor affinity [IC50=2.06nM (95% confidence interval 1.48-2.88nM)], but has higher tumor uptake (1 hours =5.65 + 1.13%ID/g, 2 hours =5.23 + 0.5%ID/g 0, 4 hour =5.39 0.84%ID/g), and the accumulation in the kidney is less than 1 hours (1 hours). 2 hours =16.84 + 3.57%ID/g; 4 hours =16.44 + 1.66%ID/g). DOTA-MSH1 receptor affinity was lowest [IC50=3.10nM (95% confidence interval 2.34-4.10nM)], with moderate tumor uptake (1 hours =2.81+1.49%ID/g; 2 hours =3.74 + 1.57%ID/g; 4 hour =4.20 0.69%ID/g) and lower kidney accumulation (1 hours + 4); 2 hours. 4 + 3.95%ID/g; 4 hours =9.90 + 1.25%ID/g). Finally, in vivo competition inhibition test on DOTA-MSH2 showed that it had good tumor specificity. DOTA-MSH4 with high receptor affinity had poor imaging effect. In the three peptide probes tested in this experiment, DOTA-MSH2 was proved to be the best melanoma PET imaging agent.
【學(xué)位授予單位】：中國(guó)協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.5

【共引文獻(xiàn)】

相關(guān)期刊論文 前5條

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相關(guān)博士學(xué)位論文 前3條

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相關(guān)碩士學(xué)位論文 前3條

1 Subhash Kumar Ghimire;[D];大連醫(yī)科大學(xué);2007年

2 李欽宗;PET-CT技術(shù)對(duì)骨質(zhì)疏松骨代謝變化的實(shí)驗(yàn)研究[D];南方醫(yī)科大學(xué);2007年

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