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Compositions and methods for loading extracellular vesicles with chemical and biological agents/molecules

A technology of drug molecules and cells, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, genetic material components, etc.

Pending Publication Date: 2020-04-10
THE RES FOUND OF STATE UNIV OF NEW YORK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Therefore, the pair can be used to block unwanted cellular communication by reprogramming exosomes to prevent and / or reverse disease progression and can be used to reprogram exosomes or load chemical and biological agents so that they can deliver other desired cargoes There is a continuing and unmet need for compositions and methods to target cells

Method used

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  • Compositions and methods for loading extracellular vesicles with chemical and biological agents/molecules
  • Compositions and methods for loading extracellular vesicles with chemical and biological agents/molecules
  • Compositions and methods for loading extracellular vesicles with chemical and biological agents/molecules

Examples

Experimental program
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Embodiment 1

[0065] figure 1 Non-limiting instructions are provided for reprogramming exosomes released from cancer cells using polynucleotides containing EXO-Code to arrest disease progression. (A) Tumor-secreted exosomes have been shown to prepare the metastatic microenvironment in distant organs. (B) Schematic depicting the conjugation of EXO-Code to therapeutic delivery to reprogram tumor-derived exosomes to arrest disease progression.

[0066] figure 2 A schematic illustrating the use of exosome biogenesis to identify biomimetic EXO-Code specifically enriched in exosomes is provided. During exosome biogenesis, ncRNA / mRNA can be loaded into exosomes during inward budding of the MVB membrane. Loading of nucleic acids can occur by: (A) passive mechanisms, (B) / (C) / (D) via active sorting mechanisms by sequence-specific sequences. (E&F) Exosomes are released into the extracellular environment by fusing with the cell membrane. (D) We adapted the process to identify the EXO-Code describ...

Embodiment 2

[0145] To assess the importance of specific nucleotides for the sorting potential of the identified EXO-Code motifs, mutant constructs containing single mutations were synthesized. In each case, C was mutated to G and vice versa, and A was mutated to U and vice versa, with the exception of consensus mutant 4, in which C was mutated to A, since this position in the motif is more commonly C, G or U instead of A. Motifs and motif hybrids spaced by 4 A nucleotides 5' upstream of the SNAIL siRNA sequence were synthesized. For the hybrid sequence (5'-AAGUGGCGUG-3') (SEQ ID NO:208), the motif is contiguous to the antisense SNAIL sequence and conjugated to the APC fluorescently labeled sense strand. For both the M8 (5'-AUCUUGUGGUC-3') (SEQ ID NO:207) and S6 (5'-AUGCGGGGGC-3') (SEQ ID NO:209) motifs, the orientation was swapped so that the active antisense The strands are conjugated fluorescent reporters. MDA or MSCs were electroporated with each construct in triplicate and washed 5...

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Abstract

Presented are RNA nucleotide sequences referred to as EXO-Codes, and longer RNA polynucleotides that contain the EXO-Codes. EXO-Codes provide RNA with the ability to a) be selectively sorted to extracellular vesicles such as exosomes, and b) deliver a variety of cargo types to program or reprogram the extracellular vesicles, and cells that receive the exosomes. Also presented are methods of makingthe EXO-Codes, modifying cells using the EXO-Codes, expression vectors encoding the EXO-Codes, and exosomes and other secreted vessices that include RNA polynucleotides that contain the EXO-Codes.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Application No. 62 / 504,941, filed May 11, 2017, and U.S. Provisional Application No. 62 / 631,060, filed February 15, 2018, the disclosures of which are hereby incorporated by reference. [0003] government fund [0004] This invention was made with government support under Grant Nos. HL126082, EB021454, and EB023262 awarded by the National Institutes of Health. The government has certain rights in this invention. technical field [0005] The present disclosure generally relates to compositions and methods for loading exosomes / extracellular vesicles with chemical and biological agents and / or for modulating cargo sorting of exosomes / extracellular vesicles. Background technique [0006] In 1 milliliter of human blood, there are approximately one billion exosomes: lipid vesicles containing proteins, miRNA and mRNA. Cells release exosomes into the extracellular environm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/11C12N15/85
CPCC12N15/10C12N15/11C12N15/63C12N15/111C12N2310/14C12N2310/141C12N2310/3519C12N2320/32A61K48/00A61K31/713A61K9/5068C12N2310/20
Inventor 朱莉安·阮斯科特·弗格森
Owner THE RES FOUND OF STATE UNIV OF NEW YORK
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