RNA chimera and application thereof

A chimera and multipurpose technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, drug combination, etc., can solve the problems of excessive immune response and lack of targeting, and achieve the release of immunosuppression, reduction of breast cancer metastasis, in situ The effect of slowing tumor growth

Inactive Publication Date: 2017-11-07
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The targeted therapy for Treg also lacks targeting; the Treg existing in the normal body will also be affected, resulting in an excessive immune response, which limits the use of this type of drug

Method used

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  • RNA chimera and application thereof
  • RNA chimera and application thereof
  • RNA chimera and application thereof

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Experimental program
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Effect test

Embodiment 1

[0026] 1. Naive CD4 + T cell infiltration is associated with regulatory T cell enrichment and poor prognosis in breast cancer

[0027] We detected the expression of CD3, CD45RA, and CD4 in 466 breast cancer sections from two independent breast cancer centers by immunohistochemical three-staining method, and analyzed naive CD4 + T cells are associated with patient prognosis). Disease-related analysis was performed using the optimal survival cut-off point calculated by X-tile statistical software. Our results show that naive CD4 + Massive infiltration of T cells (3.6 cells / field) was significantly associated with decreased disease-free survival (P value 0.016, figure 1 A). In addition, we analyzed naive CD4 in breast cancer specimens + T cells and CD3 + CD4 + Foxp3 + Correlation of regulatory T cell infiltration. Pearson correlation analysis showed that in tumor tissue, naive CD4 + The number of T cells is closely related to the number of regulatory T cells ( figure 1...

Embodiment 2

[0037] 1. Experimental method

[0038] (1) Fresh human umbilical cord blood was collected from Sun Yat-sen Memorial Hospital of Sun Yat-sen University. Hematopoietic stem cells were isolated according to the reported method, and cord blood mononuclear cells were collected by density gradient centrifugation of lymphocyte separation medium, and CD34+ hematopoietic stem cells were sorted using CD34 precursor cell separation kit magnetic beads (Miltenyi Biotec). After 2 purifications, the cell purity was detected to be greater than 95%.

[0039] (2) NOD / SCID / IL2rγnull mice were purchased from Jackson Laboratories and raised in the Animal Experiment Center of the University of Hong Kong. 3-4 weeks old female NSG mice were given a total irradiation of 200cGy, and 2x10 5 CD34+ human hematopoietic stem cells. One week after implantation of human hematopoietic stem cells, 0.1ml lentiviral vector encoding human M-CSF was injected into the tail vein of rats, with a titer of 5-10x10 8...

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Abstract

The invention provides an RNA chimera. SiRNA is specifically introduced into a T lymphocyte through the RNA chimera while other types of cells are not affected, so that the conditioned targeted therapy effect is guaranteed. The siRNA of PITPNM3 carried by a CD4 nucleic acid aptamer specifically silences expression of PITPNM3 in the T cell, the effect of CCL18 secreted by cancer related cells is blocked, the immunosuppression of breast cancer is relieved, tumor in situ grows slowly, and breast cancer metastasis is reduced.

Description

technical field [0001] The invention relates to an RNA chimera and its application. Background technique [0002] In recent years, tumor targeted therapy has been a very popular research topic, but choosing a targeting carrier with high specificity and high affinity has always been the bottleneck restricting the research of active targeted therapy, although molecules such as antibodies and peptides have been There are reports of successful researches as targeting vectors for tumor therapy, but there are insurmountable defects in the application of these compounds, and their clinical application process is greatly limited due to potential immunogenicity and preparation difficulties. At present, disease treatment and research using aptamer-mediated siRNA has developed rapidly in recent years, and more of them play a role in antiviral diseases. [0003] 1. Treatment of diseases with nucleic acid aptamers alone: ​​In December 2004, the first nucleic acid aptamer drug approved b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12N15/113A61K48/00A61K31/713A61K47/54A61P35/00
CPCC12N15/113C12N15/115C12N2310/14C12N2310/16C12N2310/32
Inventor 宋尔卫苏士成
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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