Horizontal gene transfer from macrophages to ischemic muscles upon delivery of naked DNA with Pluronic block copolymers

Abstract

Intramuscular administration of plasmid DNA (pDNA) with non-ionic Pluronic block copolymers increases gene expression in injected muscles and lymphoid organs. We studied the role of immune cells in muscle transfection upon inflammation. Local inflammation in murine hind limb ischemia model (MHLIM) drastically increased DNA, RNA and expressed protein levels in ischemic muscles injected with pDNA/Pluronic. The systemic inflammation (MHLIM or peritonitis) also increased expression of pDNA/Pluronic in the muscles. When pDNA/Pluronic was injected in ischemic muscles the reporter gene, Green Fluorescent Protein (GFP) co-localized with desmin+ muscle fibers and CD11b+ macrophages (MØs), suggesting transfection of MØs along with the muscle cells. P85 enhanced (~4 orders) transfection of MØs with pDNA in vitro. Moreover, adoptively transferred MØs were shown to pass the transgene to inflamed muscle cells in MHLIM. Using a co-culture of myotubes (MTs) and transfected MØs expressing a reporter gene under constitutive (cmv-luciferase) or muscle specific (desmin-luciferase) promoter we demonstrated that P85 enhances horizontal gene transfer from MØ to MTs. Therefore, MØs can play an important role in muscle transfection with pDNA/Pluronic during inflammation, with both inflammation and Pluronic contributing to the increased gene expression. pDNA/Pluronic has potential for therapeutic gene delivery in muscle pathologies that involve inflammation.

Targeted Delivery of siRNA Lipoplexes to Cancer Cells Using Macrophage Transient Horizontal Gene Transfer

“…The data presented focuses on siRNA, however, it is likely that other nucleic acids (i.e., pDNA, mRNA, microRNA) can also be delivered in this fashion. [29] Moreover, the finding that macrophage polarization modulates this release of siRNA inspires the design of nanoparticle carriers that can take advantage of this mechanism. This establishes a framework for macrophages as a cellular theranostic: macrophage activity can be used to study pharmacokinetics and pharmacodynamics of drug-tumor interactions as well as promote the specificity of gene delivery.…”

“…Macrophages in other contexts have been shown to transfer oligonucleotides to surrounding cells. [26,27] Our previous work has shown that macrophages can horizontally transfer proteins and DNA into neurons, [28] muscle cells, [29] and even immune cells in distal organs. [30] There are two central reasons why macrophages are ideal carriers for delivery into solid tumors.…”

Data on macrophage mediated muscle transfection upon delivery of naked plasmid DNA with block copolymers

“…MHLIM was generated as described in materials and methods of Ref. [1] and shown in a stepwise surgical procedure in Fig. 1 a.…”

“… The data contains 14 figures supporting the research article “Horizontal gene transfer from macrophages to ischemic muscles upon delivery of naked DNA with Pluronic block copolymers” [1] . The data explains the surgical procedure and histological characterization of Murine Hind Limb Ischemia.…”

Biomedical Applications of Nano-Sized Polymeric Micelles and Polyion Complexes

“…It turned out that when injected into the muscle with DNA, along with the poloxamer, which acts as a kind of "adjuvant" that activates the cells of the immune system, macrophages efficiently capture DNA, and not only transmit it to muscle cells, but can deliver it to distant organs from the injection site, in particular the lymph nodes and spleen (Gaymalov et al, 2009). It also turned out that the macrophages that captured DNA in the cell culture can "horizontally" transfer this DNA to cells of another type -muscles, neurons, cancer cells, while DNA is expressed in these new cells, even if it cannot be transcribed in the originally captured cells macrophages (Mahajan et al, 2016). Thus, transfected macrophages can act as "gene delivery systems" in other cells, including inflammation sites for various diseases.…”