Aptamer &Dendrimer Delivery of Zn Finger Nuclease &Homing Endonuclease mRNA &cDNA

适体

基本信息

批准号：
8202343
负责人：
John Joseph Rossi
金额：
$ 21.78万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-08 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8202343
关键词：
AIDS/HIV problem Acquired Immunodeficiency Syndrome Affinity Allogenic Anti-HIV Agents Binding CCR5 gene CD4 Positive T Lymphocytes CXCR4 gene Cell Culture Techniques Cells Chemokine Receptor Gene Code Complement component C5 Complementary DNA Cytoplasm DNA DNA delivery Dendrimers Development Diagnostic Disease Drug Costs Drug resistance Drug toxicity Enzymes Fingers Future Gene Targeting Genes Genetic Goals HIV HIV Envelope Protein gp120 HIV Infections HIV Receptors HIV-1 Health Hematopoietic System Hematopoietic stem cells Highly Active Antiretroviral Therapy Homing Human Image In Vitro Individual Infection Knowledge Lead Macaca nemestrina Mediating Messenger RNA Modeling Multi-Drug Resistance Mus Mutate Nucleic Acid Regulatory Sequences Nucleic Acids Patients Pharmaceutical Preparations Pharmacotherapy Population RNA Receptor Gene Research Resistance Secondary to Site-Directed Mutagenesis Small Interfering RNA Specificity Stem cell transplant Surface T-Lymphocyte Technology Testing Therapeutic Toxic effect Translations Treatment Efficacy Tropism Vertebral column Viral Virus Virus Diseases Work Zinc Fingers aptamer cell type chemokine combat combinatorial design endonuclease flexibility gene function gene therapy genetic element in vivo inhibitor/antagonist leukemia mouse model nanoparticle nonhuman primate novel novel strategies nuclease prevent programs purge receptor simian human immunodeficiency virus small molecule tool

项目摘要

HIV/AIDS continues to be a major threat to human health. The use of combinations of small molecule drugs in highly active any-retoviral therapy (HAART) to stop or thwart HIV propagation has had a major impact on delaying the progression from HIV-1 infection to the development of AIDS. Despite this progress, there are problems associated with a lifetime of anti-viral small drug therapy which include toxicity, the emergence of virus resistant to multiple drugs, and the cost of a daily, lifelong medication. The proposed studies take advantage of recent advances in the functionality of sequence specific Zn finger and homing endonucleases are capable of destroying gene function. The endonucleases are being developed in other projects of this program which will target both the chemokine co-receptor for HIV CCR5 as well as integrated proviral regulatory regions. The major challenge in using these endonucleases in a therapeutic setting is delivery of the enzymes or sequences encoding the enzymes to HIV-1 infectible cells. Since long term expression of the endonucleases could lead to secondary non-target cleavage and gene destruction, transient expression is a necessity if this approach is to be clinically useful. The proposed studies take advantage of two novel platforms for delivery of Zn finger and homing endonuclease encoding sequences into HIV (and SHIV) infectible and infected cells. The first approach uses RNA aptamers with proven ability to bind to the HIV-1 envelope expressed on the surface of HUV infected cells and internalize for functional delivery of attached siRNAs. We now will test the ability of these aptamers along with an internalizing CD4 specific aptamer to deliver backbone modified, nuclease resistant mRNAs encoding the endocucleases to uninfected and SHIV infected cells. The targets for the nucleases are the CCR5 co-receptor gene and the SHIV LTRs. As an alternative delivery approach we will test flexible TAMAM dendrimers which we have demonstrated as effective agents for siRNA deliverty in cell culture and in vivo. We will utilize this non-toxic but non-targeted delivery platform as an alternative approach for delivering backbone modified mRNAs and/or cDNAs encoding the anti-viral and anti-CCR5 endonucleases. Since both approaches have been successfully tested in vivo in a humanized mouse model for HIV infection, we will proceed to develop these agents for future application in the SHIV-pigtail macaque model utilized in other projects of this proposal. Knowledge gained from the proposed studies will result in new in vivo approaches for delivery of DNA targeting endonucleases for the treatment, and potential purging of HIV-1 infected.

艾滋病毒/艾滋病仍然是对人类健康的主要威胁。使用小分子药物的组合在高度活跃的任何归还疗法（HAART）中，停止或阻止HIV传播对延迟了从HIV-1感染到艾滋病发展的发展。尽管取得了这种进展，但仍有与抗病毒小药物疗法相关的问题，包括毒性，出现病毒对多种药物具有抗性，以及每日终身药物的成本。拟议的研究采取序列特异性锌指和归巢核酸内切酶功能方面的最新进展优势能够破坏基因功能。核酸内切酶正在其他项目中开发将针对HIV CCR5的趋化因子共受体和综合前病毒的程序监管区域。在治疗环境中使用这些测核酶的主要挑战是交付将酶编码为HIV-1感染细胞的酶或序列。由于长期表达核酸内切酶可能导致次级非目标裂解和基因破坏，瞬态表达是一个如果这种方法在临床上有用，则需要。拟议的研究利用了两个小说将锌手指和归核核酸内切酶编码序列传递到HIV（和SHIV）的平台感染和感染的细胞。第一种方法使用具有与HIV-1结合的能力的RNA适体包络在HUV感染的细胞表面表达，并内在化以递送附着的功能递送 sirnas。现在，我们将测试这些适体的能力以及内在化的CD4特异性适体传递骨链修饰的抗核酸酶耐核mRNA，编码内核酸酶至未感染和SHIV 感染细胞。核酸酶的靶标是CCR5共受体基因和SHIV LTR。作为替代交付方法，我们将测试我们已证明为细胞培养和体内siRNA deliverty的有效药物。我们将利用这种无毒但无目标的输送平台是一种替代方法，用于传递骨架修改的mRNA和/或cDNA 编码抗病毒和抗CCR5核酸内切酶。由于两种方法都已成功测试在用于艾滋病毒感染的人源化小鼠模型中，我们将继续为未来开发这些药物在本提案的其他项目中使用的Shiv-tigtail猕猴模型中的应用。知识获得了根据拟议的研究，将导致新的体内方法用于递送DNA靶向内切酶用于治疗，并可能清除HIV-1感染。