Toxicity Studies of Cardiac-Targeting Peptide Reveal a Robust Safety Profile.
Toxicity Studies of Cardiac-Targeting Peptide Reveal a Robust Safety Profile.
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DOI：
10.3390/pharmaceutics16010073
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发表时间：
2024-01-04
期刊：
影响因子：
5.4
通讯作者：
Zahid M
中科院分区：
文献类型：
--
作者：
Sahagun DA;Lopuszynski JB;Feldman KS;Pogodzinski N;Zahid M
专门针对心肌细胞的靶向治疗将为心力衰竭等常见疾病开辟新的领域。我们之前的工作使用噬菌体展示方法鉴定了一种长 12 个氨基酸的肽，该肽在静脉注射后短短 5 分钟内选择性地靶向心肌细胞，因此被称为心脏靶向肽（CTP：APHLSSQYSRT）。 CTP 已被用于向心肌细胞递送显像剂、小药物分子、光敏纳米颗粒、外泌体，甚至 miRNA。作为 CTP 作为临床上可行的心脏载体发展的自然延伸，我们现在介绍用该肽进行的毒性研究。使用 10 µM Cyanine-5.5 标记的 CTP (CTP-Cy5.5) 在人左心室肌细胞系中进行体外活力研究。完成了 CTP 的体外离子通道谱，随后对稳定转染的细胞系中的几种 GPCR 偶联受体进行了广泛的研究。通过对小鼠心脏组织进行 RT-qPCR 进一步询问 GPCR 偶联受体的阳性数据。体内研究包括单次 CTP (10 mg/Kg) 注射后的急性血压监测。进一步的体内毒性研究包括向 60 只 6 周龄野生型 CD1 雄性/雌性小鼠 (1:1) 注射 CTP (150 µg/Kg)，并在第 0、1、2、7 和 14 天用吸入 CO2 对小鼠进行安乐死，然后通过心脏穿刺采血、全血细胞计数分析、代谢分析，最后进行肝脏、肾脏和甲状腺分析研究。最后，在注射 CTP (150 µg/Kg) 之前和之后立即进行小鼠心脏 MRI，以评估心脏大小或功能的变化。与 CTP-Cy5.5 孵育 30 分钟后，人左心室心肌细胞的活力没有下降。在最高测试浓度下，除了 OPRM1 和 COX2 之外，没有观察到任何 78 个蛋白通道的显着激活或抑制，而使用 RT-qPCR 评估，这两种通道均不在小鼠心脏组织中表达。 CTP (10 mg/Kg) 注射不会导致血压变化。血细胞计数和化学分析显示没有明显的血液学、肝脏或肾脏毒性的证据。最后，CTP 注射对心脏功能、大小或质量的急性反应没有差异。我们的 CTP 研究表明体外 GPCR 相关受体没有激活或抑制。我们没有发现表明体内毒性的信号。最重要的是，心脏功能对 CTP 摄取的反应保持急剧变化。在考虑进行人体研究之前，需要使用良好的实验室实践进行进一步的研究，长期、长期地施用与所选特定货物缀合的 CTP。
Targeted delivery of therapeutics specifically to cardiomyocytes would open up new frontiers for common conditions like heart failure. Our prior work using a phage display methodology identified a 12-amino-acid-long peptide that selectively targets cardiomyocytes after an intravenous injection in as little as 5 min and was hence termed a cardiac-targeting peptide (CTP: APHLSSQYSRT). CTP has been used to deliver imaging agents, small drug molecules, photosensitizing nanoparticles, exosomes, and even miRNA to cardiomyocytes. As a natural extension to the development of CTP as a clinically viable cardiac vector, we now present toxicity studies performed with the peptide. In vitro viability studies were performed in a human left ventricular myocyte cell line with 10 µM of Cyanine-5.5-labeled CTP (CTP-Cy5.5). In vitro ion channel profiles were completed for CTP followed by extensive studies in stably transfected cell lines for several GPCR-coupled receptors. Positive data for GPCR-coupled receptors were interrogated further with RT-qPCRs performed on mouse heart tissue. In vivo studies consisted of pre- and post-blood pressure monitoring acutely after a single CTP (10 mg/Kg) injection. Further in vivo toxicity studies consisted of injecting CTP (150 µg/Kg) in 60, 6-week-old, wild-type CD1, male/female mice (1:1), with cohorts of mice euthanized on days 0, 1, 2, 7, and 14 with inhalational CO2, followed by blood collection via cardiac puncture, complete blood count analysis, metabolic profiling, and finally, liver, renal, and thyroid studies. Lastly, mouse cardiac MRI was performed immediately before and after CTP (150 µg/Kg) injection to assess changes in cardiac size or function. Human left ventricular cardiomyocytes showed no decrease in viability after a 30 min incubation with CTP-Cy5.5. No significant activation or inhibition of any of seventy-eight protein channels was observed other than OPRM1 and COX2 at the highest tested concentration, neither of which were expressed in mouse heart tissue as assessed using RT-qPCR. CTP (10 mg/Kg) injections led to no change in blood pressure. Blood counts and chemistries showed no evidence of significant hematological, hepatic, or renal toxicities. Lastly, there was no difference in cardiac function, size, or mass acutely in response to CTP injections. Our studies with CTP showed no activation or inhibition of GPCR-associated receptors in vitro. We found no signals indicative of toxicity in vivo. Most importantly, cardiac functions remained unchanged acutely in response to CTP uptake. Further studies using good laboratory practices are needed with prolonged, chronic administration of CTP conjugated to a specific cargo of choice before human studies can be contemplated.
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影响因子：
2.8
作者：
Mailhiot, Sarah E.;Thompson, Matthew A.;June, Ronald K.
通讯作者：
June, Ronald K.
DOI：
10.15171/bi.2017.17
发表时间：
2017
期刊：
BioImpacts : BI
影响因子：
--
作者：
Sabouri-Rad S;Oskuee RK;Mahmoodi A;Gholami L;Malaekeh-Nikouei B
通讯作者：
Malaekeh-Nikouei B
影响因子：
5.4
作者：
Hingorani DV;Camargo MF;Quraishi MA;Adams SR;Advani SJ
通讯作者：
Advani SJ
影响因子：
12.4
作者：
Mi, ZB;Lu, XL;Robbins, PD
通讯作者：
Robbins, PD
影响因子：
3.4
作者：
Manosroi, Jiradej;Lohcharoenkal, Warangkana;Manosroi, Aranya
通讯作者：
Manosroi, Aranya