-
细胞凋亡又称程序性细胞死亡,是多细胞生物在胚胎发育、正常组织稳态的维持与各种生理病理情况下机体清除多余细胞的重要方式。多数疾病的发生发展均与凋亡过程紧密相关,如自身免疫性疾病、神经退行性疾病(细胞凋亡发生过多)或肿瘤的发生(细胞凋亡发生过少)[1]。肿瘤经有效的治疗后会引起细胞凋亡。因此,监测细胞凋亡的过程具有重要意义。
细胞凋亡途径主要归纳为以下3种:一是线粒体凋亡途径,又称内源性凋亡途径,由B淋巴细胞瘤2超家族成员介导;二是死亡受体途径,又称外源性凋亡途径,由特异的死亡受体与相应的配体结合介导;三是内质网应激反应介导的细胞凋亡途径[2]。Caspase(天冬氨酸特异性半胱氨酸蛋白酶)家族在程序性细胞死亡过程中发挥着非常重要的作用,通常情况下Caspase以酶原的形式存在于细胞质内。各种凋亡途径最终汇聚于执行分子Caspase-3的激活,Caspase-3的激活最终会导致多聚ADP核糖聚合酶的激活,并伴随凋亡细胞形态学改变[3]。凋亡的一个早期变化是磷脂酰丝氨酸(phosphatidylserine,PS)外翻,在活细胞中PS分布于细胞膜脂质双层的内侧,一旦凋亡发生,PS便外翻至细胞脂质双层的外侧。随着PS外翻,细胞质皱缩,随后细胞皱缩,质膜出泡,细胞分裂为凋亡小体。由于在形态学与分子生物学上非常相似,细胞凋亡常与其他类型的细胞死亡混淆,如细胞自噬与坏死。因此,寻找区分凋亡与其他类型细胞死亡过程的方法具有重要意义。
传统的凋亡检测方法主要包括光学显微镜观察、原位末端标记法(TdT-mediated-dUTP nick and labeling,TUNEL)分析、流式细胞仪检测等[4],但却存在着一些固有的缺点,如均需活体取材,属于有创性的检测方式;通常只能在某一时间点观察凋亡,无法动态监测凋亡在体内的发生与发展过程。而活体内凋亡显像更有助于无创观察、理解凋亡发生的体内过程。核素凋亡显像是研究最早、最成熟的在体凋亡检测技术,原理是通过放射性核素标记化合物并与细胞凋亡过程中产生的特有靶分子结合,通过影像学手段检测细胞凋亡部分浓聚的探针分子从而达到显示凋亡的目的。本文着重从凋亡生物学过程入手,利用核医学显像剂标记技术介绍几种比较明确的细胞凋亡分子探针,为凋亡显像的临床应用提供相关的基础及指导。
放射性核素标记的凋亡显像剂的研究进展
Recent advances in apoptosis imaging using radionuclide-labeled tracers
-
摘要: 细胞凋亡存在于多种病理过程中, 包括神经系统变性疾病、缺血性损伤、自身免疫性疾病和多种肿瘤等。凋亡检测的可视化对疾病的诊断、新的治疗方法的开发与疗效评价具有重要意义。传统的凋亡检测方法包括光学显微镜观察、原位末端标记法分析、流式细胞仪检测等, 但其侵入性方式限制了之后的随访研究。而活体内凋亡显像有助于无创观察、直观了解凋亡发生的体内过程。PET与SPECT的发展, 以及新的针对靶点的放射性核素标记显像剂的合成, 使核医学进入了分子影像学的新时代。近年来, 细胞凋亡PET与SPECT显像剂的研发应用, 使活体内无创PET与SPECT检测细胞凋亡成为现实。笔者主要介绍用于在体凋亡显像的放射性标记探针及其最新研究应用进展。
-
关键词:
- 细胞凋亡 /
- 分子探针 /
- 放射性核素显像 /
- 正电子发射断层显像术 /
- 体层摄影术, 发射型计算机, 单光子
Abstract: Apoptosis or programmed cell death is an important form of cell death. Apoptosis is involved in numerous human pathological conditions, such as neurodegenerative diseases, ischemic damage, autoimmune disorders, and many types of cancer. Visualization of apoptosis is enormously beneficial in clinical diagnosis, development of new therapies, and therapeutic evaluation. The traditional methods of apoptosis detection include optical microscopy, TdT-mediated-dUTP nick end labeling analysis, and flow cytometry. However, these invasive techniques restrict the conduct of follow-up studies. Apoptosis imaging in living subjects has contributed to nondestructive observation and in understanding the biological process of apoptosis. The developments in PET and SPECT technologies, including the synthesis of targeted radionuclide tracers, led nuclear medicine into a new era of molecular imaging. The development and application of PET and SPECT as apoptosis imaging probes rendered the non-invasive detection of apoptosis in vivo a reality. This article reviewed the recent advances in apoptosis imaging using radionuclide-labeled tracers. -
[1] Kerr JF, Wyllie AH, Currie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics[J]. Br J Cancer, 1972, 26(4):239-257. [2] Green DR, Kroemer G. The pathophysiology of mitochondrial cell death[J]. Science, 2004, 305(5684):626-629. doi: 10.1126/science.1099320 [3] Levine B, Kroemer G. Autophagy in the pathogenesis of disease[J]. Cell, 2008, 132(1):27-42. [4] Hakumäki JM, Liimatainen T. Molecular imaging of apoptosis in cancer[J]. Eur J Radiol, 2005, 56(2):143-153. [5] Lahorte CM, Vanderheyden JL, Steinmetz N, et al. Apoptosis-detecting radioligands:current state of the art and future perspectives[J]. Eur J Nucl Med Mol Imaging, 2004, 31(6):887-919. doi: 10.1007/s00259-004-1555-4 [6] Lee S, Xie J, Chen X. Peptides and peptide hormones for molecular imaging and disease diagnosis[J]. Chem Rev, 2010, 110(5):3087-3111. doi: 10.1021/cr900361p [7] Gerke V, Moss SE. Annexins:from structure to function[J]. Physiol Rev, 2002, 82(2):331-371. [8] Oling F, Bergsma-Schutter W, Brisson A. Trimers, dimers of trimers, and trimers of trimers are common building blocks of annexin a5 two-dimensional crystals[J]. J Struct Biol, 2001, 133(1):55-63. doi: 10.1006/jsbi.2000.4337 [9] Kemerink GJ, Boersma HH, Thimister PW, et al. Biodistribution and dosimetry of 99mTc-BTAP-annexin-V in humans[J]. Eur J Nucl Med, 2001, 28(9):1373-1378. doi: 10.1007/s002590100578 [10] Blankenberg FG, Katsikis PD, Tait JF, et al. In vivo detection and imaging of phosphatidylserine expression during programmed cell death[J]. Proc Natl Acad Sci U S A, 1998, 95(11):6349-6354. doi: 10.1073/pnas.95.11.6349 [11] Kartachova MS, Valdés Olmos RA, Haas RL, et al. 99mTc-HYNIC-rh-annexin-V scintigraphy:visual and quantitative evaluation of early treatment-induced apoptosis to predict treatment outcome[J]. Nucl Med Commun, 2008, 29(1):39-44. [12] Yang DJ, Azhdarinia A, Wu P, et al. In vivo and in vitro measurement of apoptosis in breast cancer cells using 99mTc-EC-annexin V[J]. Cancer Biother Radiopharm, 2001, 16(1):73-83. doi: 10.1089/108497801750096087 [13] Lu C, Jiang Q, Hu M, et al. Preliminary biological evaluation of novel 99mTc-Cys-annexin A5 as a apoptosis imaging agent[J]. Molecules, 2013, 18(6):6908-6918. doi: 10.3390/molecules18066908 [14] Bauwens M, De Saint-Hubert M, Devos E, et al. Site-specific 68Ga-labeled Annexin A5 as a PET imaging agent for apoptosis[J]. Nucl Med Biol, 2011, 38(3):381-392. [15] Benali K, Louedec L, Azzouna RB, et al. Preclinical validation of 99mTc-annexin A5-128 in experimental autoimmune myocarditis and infective endocarditis: comparison with 99mTc-HYNIC-annexin A5[J/OL]. Mol Imaging, 2014, 13: 1-10[2015-07-09].http://www. ncbi. nlm. nih. gov/pubmed/?term=25431156. [16] Li X, Link JM, Stekhova S, et al. Site-specific labeling of annexin V with F-18 for apoptosis imaging[J]. Bioconjug Chem, 2008, 19(8):1684-1688. doi: 10.1021/bc800164d [17] Yagle KJ, Eary JF, Tait JF, et al. Evaluation of 18F-annexin V as a PET imaging agent in an animal model of apoptosis[J]. J Nucl Med, 2005, 46(4):658-666. [18] Murakami Y, Takamatsu H, Taki J, et al. 18F-labelled annexin V:a PET tracer for apoptosis imaging[J]. Eur J Nucl Med Mol Imaging, 2004, 31(4):469-474. doi: 10.1007/s00259-003-1378-8 [19] Wängler C, Wängler B, Lehner S, et al. A universally applicable 68Ga-labeling technique for proteins[J]. J Nucl Med, 2011, 52(4):586-591. [20] Bauwens M, De Saint-Hubert M, Devos E, et al. Site-specific 68Ga-labeled Annexin A5 as a PET imaging agent for apoptosis[J]. Nucl Med Biol, 2011, 38(3):381-392. [21] Wang F, Fang W, Zhang MR, et al. Evaluation of chemotherapy response in VX2 rabbit lung cancer with 18F-labeled C2A domain of synaptotagmin I[J]. J Nucl Med, 2011, 52(4):592-599. [22] Poulsen RH, Rasmussen JT, Ejlersen JA, et al. Pharmacokinetics of the phosphatidylserine tracers 99mTc-lactadherin and 99mTc-annexin V in pigs[J/OL]. EJNMMI Res, 2013, 3(1): 15[2015-07-09].http://www. ejnmmires. com/content/3/1/15. [23] Song S, Xiong C, Lu W, et al. Apoptosis imaging probe predicts early chemotherapy response in preclinical models:A comparative study with 18F-FDG PET[J]. J Nucl Med, 2013, 54(1):104-110. [24] Marconescu A, Thorpe PE. Coincident exposure of phosphatidylethanolamine and anionic phospholipids on the surface of irradiated cells[J]. Biochim Biophys Acta, 2008, 1778(10):2217-2224. doi: 10.1016/j.bbamem.2008.05.006 [25] Wang K, Purushotham S, Lee JY, et al. In vivo imaging of tumor apoptosis using histone H1-targeting peptide[J]. J Control Release, 2010, 148(3):283-291. doi: 10.1016/j.jconrel.2010.09.010 [26] Koulov AV, Stucker KA, Lakshmi C, et al. Detection of apoptotic cells using a synthetic fluorescent sensor for membrane surfaces that contain phosphatidylserine[J]. Cell Death Differ, 2003, 10(12):1357-1359. doi: 10.1038/sj.cdd.4401315 [27] Wyffels L, Gray BD, Barber C, et al. Synthesis and preliminary evaluation of radiolabeled bis(Zinc(II)-dipicolylamine) coordination complexes as cell death imaging agents[J]. Bioorg Med Chem, 2011, 19(11):3425-3433. doi: 10.1016/j.bmc.2011.04.029 [28] Oltmanns D, Zitzmann-Kolbe S, Mueller A, et al. Zn(II)-bis(cyclen)complexes and the imaging of apoptosis/necrosis[J]. Bioconjug Chem, 2011, 22(12):2611-2624. doi: 10.1021/bc200457b [29] Grimberg H, Levin G, Shirvan A, et al. Monitoring of tumor response to chemotherapy in vivo by a novel small-molecule detector of apoptosis[J]. Apoptosis, 2009, 14(3):257-267. doi: 10.1007/s10495-008-0293-7 [30] Reshef A, Shirvan A, Waterhouse RN, et al. Molecular imaging of neurovascular cell death in experimental cerebral stroke by PET[J]. J Nucl Med, 2008, 49(9):1520-1528. doi: 10.2967/jnumed.107.043919 [31] Höglund J, Shirvan A, Antoni G, et al. 18F-ML-10, a PET tracer for apoptosis:first human study[J]. J Nucl Med, 2011, 52(5):720-725. doi: 10.2967/jnumed.110.081786 [32] Bleackley RC, Heibein JA. Enzymatic control of apoptosis[J]. Nat Prod Rep, 2001, 18(4):431-440. doi: 10.1039/a909080k [33] Challapalli A, Kenny LM, Hallett WA, et al. 18F-ICMT-11, a caspase-3-specific PET tracer for apoptosis:biodistribution and radiation dosimetry[J]. J Nucl Med, 2013, 54(9):1551-1556. doi: 10.2967/jnumed.112.118760 [34] Zhou D, Chu W, Rothfuss J, et al. Synthesis, radiolabeling, and in vivo evaluation of an 18F-labeled isatin analog for imaging caspase-3 activation in apoptosis. Bioorg Med Chem Lett, 2006, 16(19): 5041-5046. [35] Wang F, Wang Z, Hida N, et al. A cyclic HSV1-TK reporter for real-time PET imaging of apoptosis[J]. Proc Natl Acad Sci USA, 2014, 111(14):5165-5170. doi: 10.1073/pnas.1321374111 [36] Yaghoubi SS, Gambhir SS. PET imaging of herpes simplex virus type 1 thymidine kinase(HSV1-tk) or mutant HSV1-sr39tk reporter gene expression in mice and humans using[18F]FHBG. Nat Protoc, 2006, 1(6): 3069-3075. [37] Banerji U. Heat shock protein 90 as a drug target:Some Like It Hot[J]. Clin Cancer Res, 2009, 15(1):9-14. doi: 10.1158/1078-0432.CCR-08-0132 [38] Van De Wiele C, Lahorte C, Vermeersch H, et al. Quantitative tumor apoptosis imaging using technetium-99m-HYNIC annexin V single photon emission computed tomography[J]. J Clin Oncol, 2003, 21(18):3483-3487. doi: 10.1200/JCO.2003.12.096 [39] 兰晓莉, 张永学, 何勇.凋亡显像剂99mTc-HYNIC-annexin V对肿瘤模型化疗疗效早期评价的可行性[J].中华肿瘤杂志, 2008, 30(10):737-740. doi: 10.3321/j.issn:0253-3766.2008.10.005
[40] Qin H, Zhang MR, Xie L, et al. PET imaging of apoptosis in tumor-bearing mice and rabbits after paclitaxel treatment with 18F- Labeled recombinant human His10-annexin V[J]. Am J Nucl Med Mol Imaging, 2015, 5(1):27-37. [41] Nguyen QD, Lavdas I, Gubbins J, et al. Temporal and spatial evolution of therapy-induced tumor apoptosis detected by caspase-3-selective molecular imaging[J]. Clin Cancer Res, 2013, 19(14):3914-3924. doi: 10.1158/1078-0432.CCR-12-3814 [42] Zhang Y, Stevenson GD, Barber C, et al. Imaging of rat cerebral ischemia-reperfusion injury using 99mTc-labeled duramycin[J]. Nucl Med Biol, 2013, 40(1):80-88. [43] Thimister PW, Hofstra L, Liem IH, et al. In vivo detection of cell death in the area at risk in acute myocardial infarction[J]. J Nucl Med, 2003, 44(3):391-396. [44] Lehner S, Todica A, Vanchev Y, et al. In vivo monitoring of parathyroid hormone treatment after myocardial infarction in mice with[68Ga] annexin A5 and[18F] fluorodeoxyglucose positron emission tomography[J/OL]. Mol Imaging, 2014, 13[2015-07-09]. http://www. ncbi. nlm. nih. gov/pubmed/?term=25249170. [45] 黄代娟, 兰晓莉, 张永学. 99mTc-HYNIC-Annexin V动脉粥样硬化斑块显像的实验研究[J].中华核医学杂志, 2008, 28(3):206-208. doi: 10.3760/cma.j.issn.2095-2848.2008.03.020
[46] D'Arceuil H, Rhine W, De Crespigny A, et al. 99mTc annexin V imaging of neonatal hypoxic brain injury[J]. Stroke, 2000, 31(11):2692-2700. [47] 朱羽苑, 黄钢.分子核医学显像展望:多参数分子显像时代[J].国际放射医学核医学杂志, 2010, 34(3):129-134. doi: 10.3760/cma.j.issn.1673-4114.2010.03.001
[48] Watanabe M, Hitomi M, Van Der Wee K, et al. The pros and cons of apoptosis assays for use in the study of cells, tissues, and organs[J]. Microsc Microanal, 2002, 8(5):375-391. doi: 10.1017/S1431927602010346 [49] Sugiura G, Kühn H, Sauter M, et al. Radiolabeling strategies for tumor-targeting proteinaceous drugs[J]. Molecules, 2014, 19(2):2135-2165. doi: 10.3390/molecules19022135
计量
- 文章访问数: 3328
- HTML全文浏览量: 2232
- PDF下载量: 3