Lorem Sit Amet Dolor
Researcher: Lorem Sit Amet

123 abc lane, Townsville, ZZ 00000, USA
Funding Progress: $§ / $§§§§§

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Glycomic Tumor Neoantigen Discovery in Diffuse Intrinsic Pontine Glioma (DIPG)
Informational
DIPG, Childhood (Brain Cancer)
Lay Summary

Diffuse intrinsic pontine glioma (DIPG) is a horrific childhood brain cancer. Patients die within 2 years of diagnosis. Treatments include radio- and chemotherapy, however the tumor always regrows due to resistance of the tumor cells to therapy. Understanding and tackling resistance is key to improve survival of patients with DIPG. We have shown that the tumor consists of a variety of cells, which have different characteristics. The cells which resist therapy are more aggressive and have a high potential to regrow the tumor. Killing these cells is crucial to therapeutic success.

We are proposing to characterize cell structures, specifically sugars (glycans), which are on the surface of cells. Our hypothesis is that distribution, size and quantity of these sugar molecule changes during therapy resistance and can be an important measure. We will identify these cell surface changes, which may serve as novel therapeutic targets using advanced analytical techniques.

Executive Summary

1. Scientific Merit: 200-400 children are diagnosed with DIPG yearly in the US and therapeutic options are dismal reflective of a 100% mortality rate. We have identified phenotypic heterogeneity among DIPG cells with a cancer stem-like subpopulation (CSCs), which expresses stem cell markers CD133 and aldehyde dehydrogenase (ALDH+). We have also found elevated expression of glycosidase in the ALDH+ CSC likely contributing to the self-renewal properties and cell plasticity. We are hypothesizing that therapeutic resistance as observed in DIPG patients stems predominantly from this resistant and aggressive stem-like population.

Our proposal aims to address and evaluate radio-and chemo resistance in DIPG. Specifically, we are interested in interrogating the glycome to identify glycomic tumor neoantigens. To obtain the N-glycosylation profile in DIPG as well as determine therapy induced glycome changes, we will be combining our studies with highly innovative glycomic (LC-ESI-MS, MALDI MSI) technologies for de novo target identification.

2. Feasibility

The aims of this project are entirely feasible and significant, as there is considerable interest in developing these mass spectrometry imaging based technologies for applications in cancer. They are based on the expertise of Dr. Galban’s group at the University of Michigan in developing murine models of pediatric- as well as adult glioma (Galban, Lemasson et al. 2012) (Galban, Al-Holou et al. 2017). Dr. Galban will be generating proposed DIPG orthotopic mouse models, utilize Bioluminescence as well as MR Imaging to monitor tumor growth and will perform radio- as well as chemotherapeutic treatments to investigate drug resistance mechanisms in DIPG. Dr. Galban is part of the Center for Molecular Imaging (CMI), which was established in 2001 at the University of Michigan as a shared resource aimed at providing state of the art imaging services for University researchers. This facility houses our state-of-the-art vivarium that is integrated together with the CMI.  The CMI facility houses within its Imaging Core a diverse range of imaging systems including MRI systems, Caliper IVIS bioluminescent/fluorescent in vivo optical imaging systems, a micro CT/Skyscan, a PET/CT system, a SPECT/CT system, a small animal dual axis confocal microscope and a Nikon confocal microscope among other equipment. The CMI has become a central, diverse and dynamic resource facility wherein exchange of techniques and ideas can occur, fostering productive interdisciplinary collaborations in cancer research. Facilities for acquisition, housing and care for research animals are provided by the Unit for Laboratory Animal Medicine (ULAM) on a fee for service basis.  The animals are maintained by ULAM in pressurized, climate-controlled, automatic light-cycled modern facilities in rooms adjacent to the laboratories of the applicant investigator. Furthermore, Dr. Galban has access to state of the art small animal radiation instrumentation: orthovoltage X-ray. We have expertise in harvesting brain tissue for subsequent histological analyses. Parraffin embedding as well as tissue sectioning will be performed by the Rogel Cancer Center histology core prior to shipment to Dr. Everest-Dass.

Dr. Everest-Dass has substantial expertise in glycan characterisation as well as advanced MALDI imaging techniques. Dr. Everest-Dass has established collaboration worldwide performing cutting-edge research applying MS imaging techniques. He currently works as a Research Scientist at the Institute for Glycomics, Griffith University. The proposed project directly fits into the priorities and strategies of the institute. Sufficient resources are immediately available, including facilities, materials and microscopy equipment. Griffith University uniquely provides the specialised interdisciplinary environment required to carry out this project, as there are available facilities for advanced glycoanalysis. Importantly the proposed study will be carried out in the recently acquired Bruker rapifleX MALDI TOF-TOF Tissuetyper. It is the state-of-the-art mass spectrometer for imaging biomolecules directly from tissue sections. The rapifleX is the most advanced TOF/TOF system available today and was re-designed specifically to meet the demands of molecular tissue imaging to include high-performance, high-throughput and high sensitivity. This is the first and only instrument of its kind in Australia and is localised at the Institute. In addition, the high resolution Orbitrap Fusion and the amaZon speed ion-trap mass spectrometers, both coupled to nanoLC chromatography systems, will immensely contribute to identification and characterisation of glycans as described in Aim 1.

3. Expertise of the investigators

Dr. Stefanie Galban is an Assistant Professor in the Department of Radiology and a member of the Center for Molecular Imaging (CMI) as well as the Rogel Cancer Center at the University of Michigan. Dr. Galban has a strong interest in developing therapeutic paradigms for many malignancies including DIPG. She has established orthotopic patient derived DIPG mouse models in her lab, which will be utilized for the proposed studies. Her expertise and interest in drug discovery, drug resistance mechanisms, cell signaling pathways, mouse models and imaging bring a strong interdisciplinary mix of ideas that are unparalleled.

This proposal will focus on bringing together many new approaches to further explore and advance our understanding of the pathobiology of DIPG, stemness as well as the therapeutic resistance observed in DIPG.

Stefanie’s group has obtained compelling and exciting results showing that a unique subset of cancer stem cells in DIPG exist, which likely confer resistance. The proposed studies will focus on interrogating the role of the glycome and glycoproteome in cell plasticity, stemness properties and drug resistance. Dr. Galban will generate the tumor models, perform bioluminescence as well as MR imaging to monitor tumor growth and will also perform all therapeutic treatments (radio- as well as chemotherapy (BCNU). Mice will be sacrificed at timepoints indicated above, formalin fixed and paraffin imbedded. Tissue block will be send to Dr. Arun Everest-Dass for analysis upon completion of the mouse work in Aim 1.

Dr. Arun Everest-Dass is a Research Scientist at the Institute for Glycomics, Griffith University in Australia. He has 9 years of experience with mass spectrometry detection and characterisation of glycans reflected in his impressive published work (29 peer-reviewed papers). Although, it has been known for decades that glycans in the extracellular environment are altered in cancer and significantly affect the regulation and progression of the disease. These alterations have been severely overlooked. Everest-Dass’s team has built an expertise in analyzing these structures and deciphering their function in cancer and other diseases. Recent technological advances especially in mass spectrometry from his group have increased interest in this field. One such development is that of mass spectrometry imaging of tissue sections. The project methodology and rationale are based on strong preliminary data achieved by Dr. Galban’s group and Dr. Everest-Dass. The aims of this project are entirely realistic, feasible and significant, as there is considerable interest in developing these technologies for applications in DIPG. The complementary mass spectrometry approaches are novel, unique and only available in a few labs worldwide. Dr. Everest-Dass will maintain overall responsibility for the entire project, and will direct the design and execution of all aims, including but not limited to: analysis and interpretation of data, writing and submitting of papers.

Plans for Investigator Interaction: Dr. Galban and Dr. Everest-Dass will be coordinating shipment of tissue blocks from the US to Australia upon completion of tissue collection in Aim 1. Dr. Everest-Dass will be working with his team to complete the proposed MS based techniques. Data analyses and interpretation will be performed by Dr. Everest-Dass and communicated to Dr. Galban electronically. Conference calls will be set up monthly to discuss study progress until overall completion of the project.

 

Description of Research Proposal

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Budget

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer gravida non felis non euismod. Fusce finibus aliquet consequat. Nam ac metus bibendum, iaculis purus sed, suscipit ligula. Proin et nisi libero. Mauris non urna urna. Nullam augue eros, fringilla sed mauris vitae, porta tincidunt risus. Aliquam sed tincidunt sem. Quisque lacinia quam tortor, imperdiet efficitur odio iaculis in. Sed ultricies condimentum volutpat. Vivamus dignissim faucibus porta.

Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.

Morbi orci urna, ornare non pretium eget, pulvinar eget magna. Ut consectetur efficitur varius. Fusce ac aliquet mauris, at mattis ligula. Quisque est libero, interdum id orci et, ornare luctus diam. Proin commodo lectus id accumsan blandit. Nulla eu turpis interdum, luctus ante ac, imperdiet tellus. In semper enim eu tristique aliquam.

Collaborations and Conflicts of Interest

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