Less sliding around
Harlequin Ichthyosis (HI) is an ultra-rare orphan skin disease where mutations in a lipid transporter protein result in a porous skin which provides no barrier function and seal. Neonatal lethality occurs in approximately 50 per cent of HI babies due primarily to dehydration and those that survive have a lifelong regime of frequently applying emollient creams, bathing and undergoing painful skin exfoliation to manage the disorder.
Monash researchers had recently discovered that an intrinsic inflammatory response was responsible for increasing HI disease severity by negatively impacting keratinocyte differentiation using mouse genetic models of HI. Building upon this novel finding the researchers set out to screen candidate anti-inflammatory drugs using an ex vivo whole skin culture assay which might be able to correct HI disease features as a first step in developing the first targeted therapeutic for the disorder.
This project required over 500 samples to be Histologically processed throughout the project lifecycle. As such it was imperative that all samples were handled in a uniform and concise manner ensuring high quality and optimal results that were consistent and comparable for each phase. To achieve this outcome Monash Histology Platform (MHP) performed the services of fixed tissue sample processing and undertook regular and rigorous initial tissue assessment (according to sample density, type and size) to determine the appropriate processing regime and staining methodology. Processed samples achieved from the in-house rapid tissue processor (Peloris) ensured high-quality, consistent results. For this project MHP also provided the access to infrastructure for DIY embedding and microtomy sectioning. MHP also performed optimisation procedures to determine appropriate DAB immunostaining protocols. Subsequent DAB immunostaining was performed by MHP using an automated Dako Autostainer and stained slides returned to the researcher for analysis. Antigenic retrieval for Fluorescent immunostaining was also performed using the Dako PTLink, after which the slides were returned to the researcher for completion of staining. Haematoxylin and Eosin staining (including Eosin optimization for this particular tissue) was carried out by MHP on the Leica Autostainer using a set of pre-determined modifications. Following delivery and quality acceptance of slides to the research team, MHP also provided on-line access to a complete project set of high resolution digital images of all H&E, DAB and IF stained sections. All slides were scanned using either the Aperio Brightfield AT or Aperio Fluorescent FL scanner. Access to Imagescope was also provided to allow visualisation and analysis of the resulting images for further comparative studies.
The services and equipment access provided by Monash Histology Platform contributed to the 2015 HMG publication of the initial mouse genetics based paper http://www.ncbi.nlm.nih.gov/pubmed/25209981
The Researchers were able to obtain and successfully carry out a Platform Access Grant to analyse the effects of the leading anti-inflammatory drug candidate in ex vivo whole skin culture screens. This further contributed to success in obtaining a small Monash Research Innovation Fund (MRIF) grant from the Monash Commercialisation Office to develop the preliminary findings to a point of commercialization. To date the findings have been provisionally patented with full patent to be submitted in mid-March 2016 and discussions have commenced with prospective industry partners.