Research group of Professor Peter Croucher

Research group of Professor Peter Croucher

Members of the Bone Biology Group, from left to right: Peter Croucher, Darren Lath, Holly Evans, Les Coulton, Michelle Lawson, Andy Chantry, Julia Hough, Orla Gallagher, Clive Buckle and Allan Williams (pictured below).

Peter Croucher is Professor of Bone Biology and Head of Human Metabolism at the University of Sheffield, UK. After completing post-doctoral training in the Department of Medicine at the University of Cambridge and later in the Department of Human Metabolism and Clinical Biochemistry at the University of Sheffield, Peter was awarded a five year Bennett Senior Fellowship by the Leukaemia Research Fund in 1997. Peter relocated to the Institute of Musculoskeletal Sciences at the University of Oxford as a Senior Research Fellow in 2001, and was appointed Professor of Bone Biology at the University of Sheffield in 2003.

A central research theme within Peter’s Bone Biology Group is to understand the cellular and molecular mechanisms of tumour-induced bone disease such as multiple myeloma, and to identify new therapeutic interventions. Almost 4000 people are diagnosed with multiple myeloma every year in the UK. Multiple myeloma is a cancer of the bone marrow and occurs when plasma cells, a type of white blood cell that normally produces antibodies against infection, undergo cancerous changes and start to grow uncontrollably in the bone marrow. The interactions of cancer cells with bone cells causes an imbalance in bone remodelling in favour of bone resorption, leading to bone lesions and increased risk of fracture.

Allan Williams

Dr. Alan Williams

Dr. Allan Williams is a research associate who joined the Bone Biology Group in 2007 after completing his Ph.D in Bone Biology at Imperial College London. Allan has developed a novel 3D imaging technique combining multiphoton and confocal microscopy to image individual GFP-labelled myeloma cells, bone and vasculature in an established multiple myeloma preclinical model (5T33MM). 3D imaging with resolution to detect single cells normally only permits tiny volumes of tissue to be modeled, which reduces the probability of observing rare event cells. Using tiled z-stack datasets of bone tissue from the 5T33MM-GFP model enables the Bone Biology Group to image large volumes of skeletal tissue, increasing the probability of observing rare event cells in situ.

Within the Bone Biology Group the Bone Analysis Laboratory team, led by Dr. Les Coulton and Orla Gallagher, provide a microCT (microcomputed tomography)/histomorphological bone analysis service for groups around the world. Correlation of microCT and microscopy datasets demonstrated the use of multiphoton generated secondary harmonics as a novel and valuable tool to image bone. Of particular relevance to the Bone Biology Group is the identification of the myeloma niche. Correlation of microscopy/microCT models permits precise localisation of individual cancer cells in the niche, both intra and extra-vascular, peri-vascular or enmeshed bone collagen. Typical microscopy scan parameters are 0-200 µM z axis, 1-6 mm X/Y axis, permitting identification of individual cells.

GFP-actin
Two dimensional correlation of MicroCT and 64-bit Volocity rendered microscopy murine calvarial models. A. Digital image of a mouse skull illustrating the scan area (boxed) modelled using microCT and microscopy. Frontal bone (F), parietal bone (P) and a region of sagittal suture (broken white line) are labelled for reference. B. MicroCT model of the calvarial frontal bone (F)/parietal bone (P) junction and sagittal suture (broken white line). The arrow indicates the right frontal bone/parietal bone junction along the coronal suture. C. 64-bit Volocity rendered tiled z stacks of the same location as B. Calvarial bone collagen (grey) imaged using multiphoton-generated secondary harmonics and rhodamine-dextran (red) imaged showing vasculature along the sagittal suture. The resolution permits visualisation of single cells in a large tissue volume. This is shown in the movie.

Allan says “Imaging cancer cells in bone is a technologically challenging goal yet is critical to establish the niche in which myeloma cells home, grow and form colonies. In our research, modelling different components of the niche is only made possible using the power of 64-bit Volocity software and a high specification computer. The Volocity 64-bit system enables me to render my large 3D volumes and the versatile and easy-to-use interface allows me to easily explore 3D environments to locate rare event cells”.

The three dimensional animation begins with a close-up of individual 5T33MM-GFP cells (green, imaged using multiphoton microscopy) deep in the sagittal suture vasculature, 18h after inoculation. As the model is zoomed out the 5T33MM-GFP cells fade into the distance, and after further rotation the vasculature labelled with rhodamine-dextran (red) is faded into the model. Further rotation introduces the secondary harmonic imaging of bone collagen (grey), and the curvature of the gross calvaria may be observed. Finally, the model zooms in, showing the majority of 5T33MM-GFP cells in the vasculature and one avascular 5T33MM-GFP cell enmeshed in bone. This single cell may form a colony.

Further information about research in the Bone Biology Group can be found at:
http://www.shef.ac.uk/medicine/staff/croucher.html
http://www.shef.ac.uk/medicine/research/corefacilities/bonelab

A recent publication from the Croucher lab using Volocity:

Lawson M A, Williams A J, Bos T, Vanderkerken k and Croucher P I. Localising individual myeloma cells to the myeloma ‘niche’ in bone using multiphoton microscopy. Bone, Volume 44, Supplement 1, May 2009, Page S164.