From Bug to Drug: A laboratory to pharma discovery loop for direct sourcing of antifungal peptide aptamers
Lead Research Organisation:
University of Manchester
Department Name: School of Biological Sciences
Abstract
We will use genetically engineered yeast cells to synthesise biologically useful molecules called peptide aptamers (PAs). PAs are designer proteins composed of short amino acid chains, encased within a protein scaffold. A unique property of PAs is their ability to fold into highly diverse 3D structures which are capable of binding very specifically to other proteins.
The construction of libraries of randomised aptamer sequences creates a source of molecules having a plethora of unique conformations, and able to recognise individual combinations of charge, shape and hydrophobicity. In an era of antibiotic resistance access to such a diverse array of molecular structures provides an invaluable source of new drugs.
PAs have several advantages over chemically-based medicines. As they bind very specifically to other proteins, PAs are likely to cause fewer side-effects than standard pharmaceuticals. The discovery pipeline requires a much shorter cycletime to obtain a potent active molecule and PAs exhibit modes of action which are not easily accessible via pharmaceutical screening, such as interfering with protein folding. The metabolism of peptide aptamers also yields naturally occurring amino acids, thereby limiting toxic side effects.
The challenge with PAs has been their delivery into fungal cells (PAs are, in general, unable to penetrate fungi), but by using nanoparticle-based drug delivery we can overcome this limitation.
All of these attributes are highly appealing from a therapeutic viewpoint and our intention to mine them represents a new way of working on drug discovery, as well as providing alternatives to conventional antimicrobial therapies.
The study will span the Manchester Fungal Infection Group (MFIG) and Blueberry Therapeutics. MFRG forms one of the foremost global centres for fungal allergy research and hosts a significant number of postgraduates and early career researchers. A placement at Blueberry will be incorporated to provide research experience in a commercial environment.
The construction of libraries of randomised aptamer sequences creates a source of molecules having a plethora of unique conformations, and able to recognise individual combinations of charge, shape and hydrophobicity. In an era of antibiotic resistance access to such a diverse array of molecular structures provides an invaluable source of new drugs.
PAs have several advantages over chemically-based medicines. As they bind very specifically to other proteins, PAs are likely to cause fewer side-effects than standard pharmaceuticals. The discovery pipeline requires a much shorter cycletime to obtain a potent active molecule and PAs exhibit modes of action which are not easily accessible via pharmaceutical screening, such as interfering with protein folding. The metabolism of peptide aptamers also yields naturally occurring amino acids, thereby limiting toxic side effects.
The challenge with PAs has been their delivery into fungal cells (PAs are, in general, unable to penetrate fungi), but by using nanoparticle-based drug delivery we can overcome this limitation.
All of these attributes are highly appealing from a therapeutic viewpoint and our intention to mine them represents a new way of working on drug discovery, as well as providing alternatives to conventional antimicrobial therapies.
The study will span the Manchester Fungal Infection Group (MFIG) and Blueberry Therapeutics. MFRG forms one of the foremost global centres for fungal allergy research and hosts a significant number of postgraduates and early career researchers. A placement at Blueberry will be incorporated to provide research experience in a commercial environment.
Publications
Bussink HJ
(2015)
Refining the pH response in Aspergillus nidulans: a modulatory triad involving PacX, a novel zinc binuclear cluster protein.
in Molecular microbiology
Loss O
(2017)
Mutual independence of alkaline- and calcium-mediated signalling in Aspergillus fumigatus refutes the existence of a conserved druggable signalling nexus.
in Molecular microbiology
Van Rhijn N
(2020)
Development of a marker-free mutagenesis system using CRISPR-Cas9 in the pathogenic mould Aspergillus fumigatus.
in Fungal genetics and biology : FG & B
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M016943/1 | 30/09/2015 | 29/09/2019 | |||
| 1640474 | Studentship | BB/M016943/1 | 30/09/2015 | 29/09/2019 | Bethany McCann |
| Description | Following exploitation of a regulatory paradigm in a non-pathogenic, fungal model organism, A. nidulans, a small cohort of chemical fragments have been identified that have the ability to prevent the pathogenically important pH signalling pathway of the ubiquitous mould A. fumigatus. Structural and chemical analysis of these compounds have lead to the definition of a novel structure-activity relationship associated with the compounds mechanism of action required to prevent functional signalling. Identified compounds act in combination to aid the activity existing antifungals, commonly used in the clinic to treat invasive A. fumigatus infections, as well as in combination with a drug, which when used alone is ineffective against A. fumigatus. |
| Exploitation Route | Novel antifungals are urgently required as currently rhere are a limited number of clinically available antifungals and towards those that are available resistance is on the rise. Consequently this study Identified a cohort of compounds with the ability to perturb the pathogenically important pH signaling pathway of the human fungal pathogen A. fumigatus. Furthermore through industrial collaboration this study identified and define a novel Structure-activity relationship that is essential for compound mediated perturbation of fungal growth. As the compounds, target and mechanism of action of these compounds appear to be novel, the outcomes of this research could be taken forward to further identify more potent, or clinically suitable, compounds which could be explored and taken onto clinical trials. Additionally the virulence essential pathway targted in this study is conserved in fungal pathogens of humans, plants and insects as such there is strong potential for agricultural exploitation of both the assay and screening techniques and identified compouds. |
| Sectors | Agriculture Food and Drink Healthcare Pharmaceuticals and Medical Biotechnology |
| Description | 8th AAA scholarship |
| Amount | £250 (GBP) |
| Organisation | Advances Against Aspergillosis |
| Sector | Hospitals |
| Start | 01/2018 |
| End | 02/2018 |
| Description | BBSRC Impact Acceleration Account - Relationship Incubator Application Form |
| Amount | £10,000 (GBP) |
| Organisation | University of Manchester |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 03/2020 |
| End | 08/2020 |
| Description | HFP Youth Travel Grant |
| Amount | € 800 (EUR) |
| Funding ID | ALC17-018 |
| Organisation | Federation of European Biochemical Societies (FEBS) |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 04/2017 |
| End | 05/2017 |
| Title | Genetic screening methodology for antifungal drug discovery |
| Description | A genetic screen was developed to identify compounds directly inhibiting fungal pH signalling. A methodology for high throughput screening was developed. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | A new funding application was submitted to BBSRC |
| Description | Identification and definition of SARs driving activity of compounds which inhibit pH signalling in the human fungal pathogen A. fumigatus. |
| Organisation | MedChemica Ltd |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | We developed a genetic screen for inhibitors of pH signalling |
| Collaborator Contribution | In silico predictions of active motifs. Identification and sourcing of chemical analogs for pinpointing structure-activity relationships. |
| Impact | A new funding application has been written and submitted to BBSRC |
| Start Year | 2019 |
| Description | Identification and definition of an SAR required for the activity of compounds with novel antifungal activity against the pH signalling pathway of the human fungal pathogen A. fumigatus. |
| Organisation | Blueberry Therapeutics |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Development and optimisation of assay used to screen compound libraries. intial library screening and compound identification. Completion of wet lab research. |
| Collaborator Contribution | Payment of a medicinal chemist to permit the further identification of structural analogues, and eventual SAR definition following screening. Provision of lab space, to complete screening assays. Time and support from experts in the field of novel antifungal drug discovery. |
| Impact | Identification and definition of an SAR, essential for compound mediated fungal perturbation. Identification of compounds with antifungal activity. Collaboration partnership with MedChemica. All above points have resulted in a collaborative grant submitted to the BBSRC responsive mode call (30/09/2020). |
| Start Year | 2015 |
| Description | Outbreak: Killer Fungus |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Public/other audiences |
| Results and Impact | Outbreak: Killer fungus. a Roleplay evernt based on an outbreak of lethal fungal diseases. participants needed to identify and treat the pathogen. additionally a series of stalls inclyding educational (microscopy, cells etc) and recreational activities (computer games, colouring, playdoh). |
| Year(s) Of Engagement Activity | 2018 |
| Description | Science Spectacular 2016 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | Science Spectacular as part of Manchester Science week. a Stall at the event entitled: 'Fungi: Friend or Foe'. addtionally a balloon release entitled the great spore race |
| Year(s) Of Engagement Activity | 2016 |