Study of immune responses to therapeutic inhaled nanoparticles by resident alveolar macrophages
Lead Research Organisation:
University of Manchester
Department Name: School of Biological Sciences
Abstract
Project Description: (maximum of 4,000 characters)
Please make sure this description clearly indicates how the project sits within the BBSRC remit, how it enables new ways of working and how it aligns with the DTP themes (World Class Underpinning Biosciences, Industrial Biotechnology and Bioenergy or Agriculture and Food Security). If you have been awarded an in vivo skills supplement, please outline the in vivo skills the student will learn during the project.
Inhaled nanoparticles are currently under active development by the pharmaceutical industry as a promising avenue for drug delivery. The technology is in its infancy and there are safety concerns, partly caused by a limited understanding of immunogenic responses to inhaled synthetic nanoparticles (NPs). NPs, defined as <0.1um in size, offer a large surface area:volume ratio, which can be used to enhance the efficiency of drug delivery. The composition of NPs varies: they can comprise natural biological materials (eg lipids, dextran) or synthetic products (eg polymers). We (TH) have previously examined the consequences of inhaled graphene particles on lung immunity and revealed their uptake by alveolar macrophages, the formation of granulomas and their alteration of macrophage reactivity. This reaction depended on the formulation and size of administered nanoparticles. We also have the expertise to test the influence of nanoparticles on immunity and pathology to respiratory allergens, viruses and bacteria in murine models.
In collaboration with MedImmune, Prof Lu's laboratory have developed a simple desktop flow device for the preparation of NPs using charged lipids, PEG, cholesterol and nucleic acid components. Particles fall into the 150-200nm size range and fundamental properties, such as charge and nucleic acid content, can be manipulated. Prior to their introduction for therapeutic use, it would be valuable to evaluate the influence of these parameters on the innate immune response in the lung, particularly alveolar macrophages.
The aims of the project are:
1. To fabricate a range of NPs with different biophysical properties (eg size, charge) by manipulation of composition using the flow device in Prof Lu's laboratory.
2. To prepare NPs with different nucleic acid composition (eg C/G rich versus A/T rich).
3. To compare the binding and uptake of NPs of different composition using membrane fluorescent probes- a variety are available, including a series with BODIPY dyes incorporated at different positions within the aliphatic lipid chain (Molecular Probes); Nucleic acid fluorescent labels (labelling kits are available, capable of attaching a variety of fluorophores eg the Alexa series).
FACS analysis will be used to study binding and uptake into tissue resident macrophages. These studies will be complemented by confocal microscopy, with a study of subcellular localization (combined with the use of appropriate subcellular markers). Specific small molecule inhibitors of antigen processing will be used to examine routes of uptake (eg endocytosis) and examine how this varies with NP size, homogeneity of preparation and other variables. Phenotypic and functional alterations in airway macrophage activity will be examined, including production of cytokines and chemokines (TNFa, IL-6, IFN, IL-29). Transcriptional profiling by NGS and quantitative RT-PCR will be used to examine changes in gene expression in response to uptake of NPs of different composition.
Please make sure this description clearly indicates how the project sits within the BBSRC remit, how it enables new ways of working and how it aligns with the DTP themes (World Class Underpinning Biosciences, Industrial Biotechnology and Bioenergy or Agriculture and Food Security). If you have been awarded an in vivo skills supplement, please outline the in vivo skills the student will learn during the project.
Inhaled nanoparticles are currently under active development by the pharmaceutical industry as a promising avenue for drug delivery. The technology is in its infancy and there are safety concerns, partly caused by a limited understanding of immunogenic responses to inhaled synthetic nanoparticles (NPs). NPs, defined as <0.1um in size, offer a large surface area:volume ratio, which can be used to enhance the efficiency of drug delivery. The composition of NPs varies: they can comprise natural biological materials (eg lipids, dextran) or synthetic products (eg polymers). We (TH) have previously examined the consequences of inhaled graphene particles on lung immunity and revealed their uptake by alveolar macrophages, the formation of granulomas and their alteration of macrophage reactivity. This reaction depended on the formulation and size of administered nanoparticles. We also have the expertise to test the influence of nanoparticles on immunity and pathology to respiratory allergens, viruses and bacteria in murine models.
In collaboration with MedImmune, Prof Lu's laboratory have developed a simple desktop flow device for the preparation of NPs using charged lipids, PEG, cholesterol and nucleic acid components. Particles fall into the 150-200nm size range and fundamental properties, such as charge and nucleic acid content, can be manipulated. Prior to their introduction for therapeutic use, it would be valuable to evaluate the influence of these parameters on the innate immune response in the lung, particularly alveolar macrophages.
The aims of the project are:
1. To fabricate a range of NPs with different biophysical properties (eg size, charge) by manipulation of composition using the flow device in Prof Lu's laboratory.
2. To prepare NPs with different nucleic acid composition (eg C/G rich versus A/T rich).
3. To compare the binding and uptake of NPs of different composition using membrane fluorescent probes- a variety are available, including a series with BODIPY dyes incorporated at different positions within the aliphatic lipid chain (Molecular Probes); Nucleic acid fluorescent labels (labelling kits are available, capable of attaching a variety of fluorophores eg the Alexa series).
FACS analysis will be used to study binding and uptake into tissue resident macrophages. These studies will be complemented by confocal microscopy, with a study of subcellular localization (combined with the use of appropriate subcellular markers). Specific small molecule inhibitors of antigen processing will be used to examine routes of uptake (eg endocytosis) and examine how this varies with NP size, homogeneity of preparation and other variables. Phenotypic and functional alterations in airway macrophage activity will be examined, including production of cytokines and chemokines (TNFa, IL-6, IFN, IL-29). Transcriptional profiling by NGS and quantitative RT-PCR will be used to examine changes in gene expression in response to uptake of NPs of different composition.
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/T508548/1 | 01/10/2019 | 28/09/2024 | |||
| 2284733 | Studentship | BB/T508548/1 | 01/10/2019 | 30/09/2020 |