As part of a series of a small grants program to stimulate translational research, the Oncological Sciences Center released five separate calls for proposals for seed funding innovative, interdisciplinary projects. The purpose of the program was to stimulate larger proposals to cancer funding agencies. Each call for proposals was based on presentations and discussions at OSC sponsored workshops.
Calls for proposals were made in the following areas in interest to the OSC:
Metobolite Profiling for Early Cancer Detection
Principal Investigators: M. Daniel Raftery, PhD, Department of Chemistry
Graham Cooks , PhD, Department of Chemistry
Abstract
This proposal will seek to develop early detection methods for cancer based on metabolite profiling. Recent results have shown that measurable changes in the concentration of metabolites present in biofluids such as serum or urine can be used to develop remarkably predictive early diagnoses of ovarian and colon cancers as well as heart disease and Alzheimer's. The Raftery and Cooks laboratories have developed advanced methods in nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry to provide sensitive and fast methods to analyze biofluid samples and to distinguish "diseased" and "healthy" populations. In the present study the newly developed method of desorption electrospray ionization (DESI) mass spectrometry will be combined with nuclear magnetic resonance (NMR) spectroscopy and principal component analysis (PCA) to shortlist compounds with potential for biomarker screening which are responsible for significant differences between control samples and diseased samples. The approach will combine high resolution spectroscopy with multivariate statistical analysis. In a recent study, we have shown that NMR and MS can be combined to differentiate mice with lung cancer from control mice based on changes in up to 80 metabolites. An important corollary of the approach is that a number of these metabolite-based "biomarkers" point to changes in metabolism, potentially providing insight into the development of the disease.
The PI's are part of an NIH-funded Center on Tools for Differential Metabolomics at Purdue, and have as a result of this funding developed the tools necessary to carry out the proposed work. Funding from the OSC will go far to help develop the preliminary data to apply for the funding from NCI for metabolite profiling. One of the PIs recently attended an NCI-sponsored workshop on metabolite profiling, and it is clear that statistically valid data from a sizable population of patients will be necessary to obtain funding. The methods that we have developed are sensitive and take under 5 min in analysis time, so they could be adopted by clinics.
Bioconjugated Gold Nanorods for In Vivo Detection and Hyperthermia of Turmor Cells
Principal Investigators: Alexander J Wei, PhD, Department of Chemistry
Ji-Xin Cheng, PhD, Department of Biomedical Engineering
Abstract
We propose to apply a robust method for conjugating bioactive ligands onto gold nanorods, for in vivo imaging of their delivery to tumor cells based on two-photon luminescence (TPL) and subsequent optical hyperthermia as an antitumor therapy. We will functionalize nanorods with pteroate ligands for targeted delivery to tumor cells which overexpress the folate receptor. The pteroate ligands will be attached by dithiocarbamate (DTC) ligation, a simple but powerful surface functionalization method recently developed by one of us. DTCs are formed by the mild condensation of alkylamines with CS2 to produce a chelating ligand with robust adsorption onto metal and inorganic surfaces. The DTC ligands are resistant to displacement by surface exchange with biogenic thiols, and can thus be used without concern for degradation in biological fluids. Cell-selective delivery of functionalized nanorods will first be demonstrated in vitro by detection and hyperthermia of KB cells mixed with non-cancerous cells, and also in vivo by detection of early-stage malignant growth using a mouse model.
Nanoscale Arrary Platform via Confocal Raman Based Multiplexing f
or Alternative Splice profiling of BRCA1
Principal Investigator: Joseph Irudayaraj , PhD, Department of Agricultural and Biological Engineering
Abstract
We will elucidate Alternative Splice (AS) profiles in nontumorigenic Human Breast epithelial Cell (HBEC) lines and compare these profiles to those obtained from tumorigenic cell lines to evaluate whether BRCA1 AS variants are specific to pathologic conditions (e.g., a malignant phenotype). Since AS profiling requires the detection of a large number of AS variants in the mRNA pool of the target gene, conventional microarray methods are not applicable because, only four predominant AS variants could be monitored. The Confocal Raman spectroscopy/imaging approach, offers several orders of magnification due to the resonance and enhancement effect and distinct vibrational mode peaks in Inm resolution which is much better that the resolution of the fluorescent bands (300-400 nm wide) or even quantum dots (40-50 nm wide peaks). Considering the choice or tags available (over 1000), practically an unlimited number of tagged AS variants could be explored, thus providing a detailed gene map. Specific aims of the Confocal Raman based multiplex AS profiling approach are to: (1) synthesize Raman-labeled oligonucleotide probes and study the magnification due to resonance and surface enhancement for five selected raman tags (fluorescent and nonfluorescent tags), (2) fabricate ~100, 50, and 1um diameter spot size (10 x 10) arrays and develop a Confocal raman strategy using appropriate laser excitations corresponding to the labels used to monitor up to five different hybridizations in one array spot, and (3) demonstrate the multiplex concept to effectively elucidate AS patterns of the BRCA1 gene from two different cell types. Long-term goal is to develop a spectroscopic procedure with data analysis and image processing capability to monitor up to 10 different biological interactions in a 1 um diameter spot thus providing unparalled multiplexing while affording nanoscale spatial sensitivity.
Live Cell and In Vivo Imaging Sstudies of Anti-Cancer Drug Delievery
Principal Investigators: Ji-Xin Cheng, PhD, Department of Biomedical Engineering
Kinam Park, PhD, Department of Industrial & Physical Pharmacy
Abstract
Development of multi-functional nanoscale drug carriers holds great promises for tumor imaging and therapy. Despite limited successes reported in each case, the mechanisms that govern the delivery of these nanocarriers are poorly understood. To provide a solid foundation for rational nanomedicine design, we propose to study the mechanisms of delivery of various nanoscale vehicles into live cells and in vivo by using molecular imaging and advanced biophysical tools. We will focus on copolymer micelles in the proposed research. Effects of the micelle size and surface ligand density on the cellular uptake efficiency and kinetics will be examined. The vehicles' endocytic pathways, intracellular trafficking, and the spatial-temporal profile of intracellular drug release will be directly visualized. Finally, in vivo optical imaging methods will be developed to visualize blood circulation and tumor targeting of the micelles intravenously injected into a mouse tumor model. The knowledge derived from our study will be essential in the design of nanoscale drug vehicles and will lead to discoveries of better delivery systems.
Identification of Cancer Drug Targets by an Engineered Dendritic
Nanomolecule and Mass Spectrometry
Principal Investigators: Weiguo Andy Tao, PhD, Department of Biochemistry
Philip S. Low, PhD, Department of Chemistry
James F. Leary, PhD, Department of Basic Medical Sciences
Abstract
Identifying drug targets is pivotal to improving new drug design, to finding potential molecular targets for therapy, and to understanding molecular mechanisms for disease treatment. New experimental strategies, in particular interdisciplinary approaches, are proposed to efficiently exploit genomic level data, highlighting chemical and proteomic approaches that aim to increase the quality of information obtained from high throughput experiments such as mass spectrometry. We seek to develop a new class of nanoprobes based on polyamidoamine dendrimers coupled with mass spectrometry to identify drug/ligand targets. Using functionalized dendrimers essentially as drug carriers, we will deliver drugs into living cells and in vivo and allow them to interact with potential intracellular targets. The dendrimers will be multi-functionalized with cancer drugs/pro-drugs, folic acid to enable dendrimer targeting and uptake by cancer cells, a reporter tag (e.g., fluorescence tag) for cell sorting, and a grip for the isolation of drug-interacting proteins. Cells incorporated with dendrimers will be selected based on the fluorescence signal, lysated, and proteins bound to dendrimers will be isolated for sequencing identification using micro-capillary liquid chromatography tandem mass spectrometry.
Multilayered Nanoparticles for Delivery of Anti-Cancer Agents
Principal Investigators: James F. Leary, PhD, Department of Basic Medical Sciences
Donald E. Bergstrom, PhD, Department of Medicinal Chemistry & Molecular Pharmacology
Deborah W. Knapp, Ph.D, DVM, Department of Veterinary Clinical Sciences
Abstract
There are many potentially effective anti-cancer agents that are of limited value due to (1) loss of activity in the bloodstream or being filtered out of the body before reaching their target cells, (2) inability to target appropriate cells which are, by definition, rare in the in-vivo situation, and (3) unacceptable bystander effects on other normal cell subpopulations caused by mistargeting. Multilayered nanoparticle (NP) drug delivery systems to be produced in this research will try to address all three problems. First, anti-cancer drugs can be shielded against molecules in the bloodstream that may inactivate the encapsulated drug. The NP can also enhance effective lifetime in the bloodstream by preventing immediate elimination and excretion, thereby significantly enhancing the pharmacokinetics. Second, the ability to target rare cells is a very difficult problem that the PI has worked on for more than 25 years and has developed many effective strategies. The targeting must be multi-step and use sophisticated Boolean molecular targeting strategies to reach levels of targeting accuracy with NPs as the PI has previously demonstrated with molecular labeling (Leary, 1994; Leary, 2000; Leary 2005). Third, as an extension of the previous argument, these targeting strategies must be designed to minimize bystander mistargeting. One way this can be accomplished is to first use a series of molecular targeting strategies involving antibodies, aptamers, and amino acid intracellular localization strategies, followed by use of intracellular molecular biosensors to error-check against false targeting. Such a strategy requires use of multilayered nanosystems such that each layer strips off under desired conditions to reveal the underlying layer which guides the next step. This strategy has been described previously by the PI in a simpler system and is the focus of a pendingUS patent application (Prow et al., 2004; Prow et al., 2005; Leary & Prow, 2005). The proposed NP research will target prostate cancer, a disease which kills >30,000 men yearly in the USA .
The overall research involves the interdisciplinary collaboration of: (1) Dr. Donald Bergstrom, a medicinal chemist who will work out a number of required steps in the chemistry of multilayered NP design, production and testing. (2) Dr. Leary for overall design of the nanosystems, targeting strategies, molecular biosensors and therapeutic genes, and (3) Dr. Deborah Knapp for in-vivo use of these nanosystems in animals to initially study toxicity, biodistribution and targeting problems, and longer term to assess the effectiveness of these nanomedicine approaches in diseased animals.
Changes in Nutrition and Lifestyle Behaviors in Response to Health Promotion in the Workplace: Support for Dissemination and Implementation Research in Cancer Prevention and Treatment
Principal Investigator: Stacey L. Mobley, PhD, RD, CNSD, Department of Foods & Nutrition
Abstract
According to the Harvard Report on Cancer Prevention, approximately 70% of all causes of cancer in the United States can be contributed to lifestyle behaviors (tobacco use, sedentary lifestyle, alcohol, sun exposure) and adult diet/obesity (diets high in animal fat and low in fruits and vegetables). The purpose of this study is to demonstrate improvements in cancer risks and clinical outcomes in response to changes in nutrition and lifestyle behaviors in adults. Approximately 11,000 university employees, spouses, and retirees from the Purdue Health Improvement Initiative will be asked to participate. Participants will be asked to complete a health risk appraisal form, wellness screening (height, weight, blood lipids and glucose, waist circumference measurements), and diet history questionnaire at baseline, 12 months, 24 months, and 36 months. A subpopulation of moderate and high health risk participants will receive behavioral counseling for nutrition/lifestyle changes via telephone for up to one year. Outcomes will include disease diagnosis, health risk appraisal scores, height, weight, waist circumference, blood pressure, blood lipids and glucose, screening tests (if available), and nutrient intake levels. Multiple linear regressions models will be used to investigate relationships between nutrition, lifestyle behavior, and clinical outcomes. Analysis of covariance with repeated-measures will be used to compare the outcomes between low, moderate and high health risk individuals. The results of this study will generate an enormous database for future outcomes research studies relative to cancer-relevant behaviors including diet, lifestyle, and exercise. It will also provide a foundation for translational research such as establishing dietary and behavioral guidelines for clinicians or relating disease outcomes to genetic-environmental interactions, thereby leading to individualized diets based on genetic and behavioral profiles.
Study of Scottish Terriers to Develop Strategies to Prevent Human Invasive Urinary Bladder Cancer
Principal Investigators: Deborah W. Knapp, DVM, MS, Dipl. ACVIM, Department of Veterinary Clinical Sciences
Jose A. Ramos-Vara, DVM, PhD, Dipl. ECVP, Department of Veterinary Pathobiology
Deepika Dhawan, PhD, Department of Basic Medical Sciences
Abstract
Invasive urinary bladder cancer (invasive transitional cell carcinoma, InvTCC) causes >14,000 deaths yearly in the US . Because therapy of advanced InvTCC is met with dismal results, it is imperative to develop strategies to attack InvTCC earlier, ideally by defining its causes and preventing it from forming. Smoking is a major cause of InvTCC, but 50% of patients with InvTCC have no known risk factors. Critical gene-environment interactions are thought to lead to InvTCC, but these are largely undefined. It is very difficult to determine genetic factors in humans because of confounding variables. We have the unique opportunity to define genetic factors and crucial gene-environment factors in Scottish terrier dogs (STs) and to then translate these findings into humans. InvTCC in STs mimics human InvTCC in histopathology and biologic behavior. The 18-fold increased risk for InvTCC in STs (compared to other breeds and mixed breed dogs) indicates a strong genetic component to the disease. Environmental factors (including lawn herbicides) further increase the risk, and specific dietary factors reduce the risk of InvTCC in STs. STs appear to be an excellent model to study gene and gene-environment interactions and prevention strategies in InvTCC. Our goals are to identify genetic risk factors for InvTCC in STs, to then determine the role of these factors in human InvTCC risk, and to use this information to develop strategies to prevent the formation and/or progression of this deadly disease. We will apply for major extramural funding to pursue this work (including an application to NIH PA-05-049 , "Animal Models of NIDDK-Relevant Diseases"). To submit a competitive NIH application, however, two tasks must first be accomplished: (1) characterize InvTCC in STs at the molecular level to further confirm it is a relevant model, and (2) generate preliminary findings of a genome scan in STs. The proposed work is aimed at these tasks. Briefly, with pet owner consent, InvTCC tissues samples will be collected from 20 STs who are undergoing cystoscopy or surgery as indicated for their treatment at the Purdue University Veterinary Teaching Hospital. PCR and Western blotting (Dhawan design, Knapp lab) and immunohistochemistry (Ramos-Vara lab) will be used to analyze genes of known importance in human InvTCC (e.g. p53, Rb, EphA2, cox-2, E-cadherin, bFGF, caspase-3, survivin, p16). For the genome scan, blood samples will be collected from STs with InvTCC and age-matched STs without InvTCC at Purdue University and at other veterinary hospitals. Dr. Ostrander will analyze these samples using an automated 500-microsatellite based genome scan, and will identify areas of the genome for further study. Dr. Glickman will be involved in study design, and will serve as our link to Banfield Pet Hospitals and their computerized data base. This link puts us in contact with large numbers of STs (25,000 ST visits, 2002-05).
Targeting connexin 43 to prevent breast cancer development from early pre-neoplastic stages
Principal Investigators: Sophie A. Lelièvre, DVM, Ph.D., Department of Basic Medical Sciences
Rebecca Doerge, Professor of Statistics and Agronomy, Department of Statistics
Abstract
This proposal focuses on characterizing the status and role of connexin-43 (Cx-43), a gap junction protein, in early breast cancer. The authors have observed the loss of Cx-43 expression in cells with altered cellular architecture present in early stages of breast cancer lesions. There is also evidence that loss of Cx-43 correlates with a loss of control of proliferation of the cells. This leads them to the hypothesis that loss of Cx-43 is an early step in the progression of tumor development and understanding its function could lead to prevention approaches. The two aims focus on different aspects of the role of Cx-43 in early breast cancer. The first aim plans to look at breast biopsies and try to correlate the loss of apical polarity of cells in tumors and in cells near tumors with the expression of different markers of proliferation and Cx-43 status. The second aim looks at the effect of reintroducing Cx-43 into different cell lines and observing the effects on proliferation and cellular architecture.
Identification of the Invasive/Metastatic Subpopulations of Breast Cancer Pregenitor Cells
Principal Investigators: Joseph Irudayaraj, Purdue University
Harikrishna Nakshatri, Indiana University School of Medicine
Abstract
Recent studies have identified a subpopulation of breast cancer cells lacking the expression of lineage specific markers and CD24 while expressing CD44 (CD44+CD24-Lin- cells) as tumorigenic progenitor cells (Al-Hajj et al., 2003). However, the relationship between progenitor status and metastatic potential is not clear. Cancer-specific stem/progenitor cells have been described for a variety of hematological malignancies as well as solid tumors (Jordan and Guzman, 2004; Singh et al. 2004). Such tumorigenic cells were distinguished from their non-tumorigenic counterparts by a specific cell surface marker profile, CD44+/CD24-/Lineage-. However, the metastasis potential and the sensitivity of these cells to chemotherapy and the effect of therapy on the syrface markers are not known or understood.
The recent gene expression profiling studies have challenged the long held view that metastatic cells are rare and arise during later stages of tumor progression due to progressive accumulation of mutations and that most primary tumors have a fraction of cells with "metastatic phenotype" (Panter and Brankenhoff, 2004; van't Veer, L. J. et al., 2002). Cells with such "metastatic phenotype" are yet to be identified and their relationship to stem cell origin of cancer is yet to be established.
Current chemotherapy shrinks but does not eliminate metastatic tumors possibly leaving behind cancer stem cells, with regenerative potential. To our knowledge, this is the first study on the identification of metastatic cancer cell subpopulation based on gene and surface marker kinetics in relation to the stemness at a single cell level. Most investigations dealing with metastasis so far utilized established cell lines because of technical difficulties with primary tumor samples. We have already established a model system to study this complex biological system using primary breast cancer cells. Highly sensitive assays using established microarray and cytometry to supplement a novel nanorod-based biosensor approach will help track the evolutionary state of the disease very early in its transformation and monitor the state of biomarkers during the course of therapy administration.
The development of single particle diagnostic assays for identification of the sparse cells will be a significant prognostic step in diagnosis and subsequent assessment of the efficacy of therapy.
Probing the Cytotoxicity of Nanomedical Systems for Cancer Therapy
Principal Investigators: James F. Leary, Ph.D. SVM Endowed Professor of Nanomedicine, Professor of Basic Medical Sciences and Biomedical Engineering, Oncological Sciences Center, Purdue Cancer Center, Purdue University
James E. Klaunig, Ph.D. Robert B. Forney Endowed Professor of Toxicology, Director, Center for Environmental Health, Associate Director IU Cancer Center, Indiana University
Abstract
Nanomaterials are being used to construct nanodelivery systems for drugs and genes to combat a wide variety of cancers. Many of these nanomaterials are hydrophobic necessitating the addition of surface molecules to promote water solubility. Some of these molecules also act as biocoatings to increase biocompatibility and to reduce toxicity. The problem is that these biocoatings can be stripped off differentially by different cell types. Light and oxidation can also cause breakdown of nanostructures, typically of lower toxicity, generating elemental forms which typically are of higher toxicity. For in vivo use of nanomaterials in nanomedicine, the breakdown of biocoatings and nanostructures is hard to predict. Complicating this process of toxicity prediction are the body's response to nanomaterials in terms of (1) clearance through the kidney, (2) breakdown of the nanomaterials into smaller components, perhaps without biocoatings, and (3) temporary sequestering of nanomaterials which reduces short-term toxicity but slows natural clearance from the body. Further complicating this picture of predicting in-vivo toxicity is the fact that nanodelivery of drugs represents a fundamental paradigm shift in terms of traditional tests of in-vivo toxicity. Since the very point of using nanodelivery systems for drugs is to prevent free diffusion of drug throughout the body and reduce toxic exposure to non-targeted bystander cells, it makes little sense to test these new nanomaterials at the kinds of concentrations typically used for testing the toxicity of traditional drugs. As cited in a recent review: "In general, there are discrepancies in the current literature regarding the toxicity of QDs (Quantum Dots) that can be attributed to several factors: the lack of toxicology-based studies, the variety of QD dosage/exposure concentrations reported in the literature, and the widely varying physicochemical properties of individual QDs. Studies specifically designed for toxicologic assessment (e.g., dose, duration, frequency of exposure, mechanisms of action) are few. Many of the studies from which QD toxicity information is derived and that have been cited in reference to QD toxicity were performed by nanotechnology researchers rather than toxicologists or health scientists" (Hardman, 2006). How should new nanotoxicity standards and predictions by made? That is the central question driving the proposed collaboration. We wish to make some initial studies that will point the way toward a new standard that can be developed in a longer range research program linking a bionanotechnologist recently recruited to Purdue University (Dr. Leary) with medical toxicologist (Dr. Klaunig) at Indiana University (IUPUI). We will start with QD cores that have some clear toxicity issues, and then extend the studies to ferric oxide toxicity which are much less and more subtle.
Delivery of nanoshells to hypoxic areas of breast tumors using a Trojan Horse
Principal Investigators: Susan E. Clare, M.D., Ph.D.,
Assistant Professor
Division of Breast Surgical Oncology
Department of Surgery, Indiana University School of Medicine
Rashid Bashir, Ph.D.,
Professor, School of Electrical and Computer Engineering,
Professor, Weldon School of Biomedical Engineering & School of Mechanical Engineering (by Courtesy),
Purdue University Faculty Scholar, Purdue University
Abstract
After receiving the best chemotherapy regime yet to be developed for the treatment of breast cancer, doxorubicin and cyclophosphamide followed by paclitaxel and Herceptin, 15% of patients will suffer a recurrence by 4 years [1]. Why? There are four explanations which are not mutually exclusive: 1.) innate resistance; 2. acquired resistance; 3.) inadequate delivery; 4.) quiescence, i.e., cells are in G0 during therapy. In this proposal we will focus on the inefficient delivery of otherwise efficacious treatments and will propose a solution to this problem.
How do we know that there is inefficient or insufficient delivery? We make this supposition because it is well documented that the delivery of oxygen and nutrients to tumors is inadequate. Frequently, the center of a tumor, the area furthest from the vascular supply, becomes hypoxic, hypoglycemic and cell death is extensive. In addition to areas of frank necrosis, microelectrode measurements of oxygen tension in breast tumors have shown that 40% of breast tumors examined contained areas of oxygen tension in the range of 0-2.5 mm Hg (normal tissues range from 20-66 mm Hg) [2]. These areas of hypoxia can also be observed in tissue sections by immunolabeling of the reductively activated hypoxic-specific marker pimonidazole [3].
Response to hypoxia
Macrophages have been shown to infiltrate breast tumors; they are directed there. Tumor cells secrete elevated levels of CC chemokines, a group of chemokines with adjacent cysteines that are chemoattractants for lymphocytes, monocytes, eosinophils, basophils but not neutrophils. As areas of hypoxia/necrosis develop and expand, monocyte/tumor associated macrophage chemoattractants such as VEGF, endothelins, EMAP II, and other yet to be identified factors are released, which perhaps are the consequence of the binding of hypoxia inducible transcription factors such as HIF-1a and HIF-2 and possibly ATF-4, Ets-1, and/or NF?B [3]. Together or individually, these factors attract monocytes into tumors and then direct the tumor activated macrophages toward areas of hypoxia/necrosis along a chemoattractant gradient. 2
We propose to utilize macrophages as a Trojan Horse to deliver anti-tumor agents to areas of hypoxia, regions of tumors otherwise inaccessible to conventional vascular delivered therapies. The anti-tumor agents we propose to use are gold nanoshells. We will take advantage of two phenomena of gold/silica nanoshells: 1.) They can be produced to macrophagesSW CNTsORNanoshellsMovement along a Chemo-attractant gradientTumorh? either preferentially absorb or scatter light by varying the size of the particle relative to the wavelength of the light at their optical resonance [4]. The absorbed light is converted to thermal energy which theoretically produces a temperature at the particle as high as 800° Kelvin [5] and experimentally has been shown to increase the temperature in the tissue in which the nanoshells are embedded by over 30°C [6]. 2.) These nanoshells can be "tuned" such that they absorb in the near infrared, a region of the spectrum where optical absorption of the tissue is minimal and penetration by the radiation optimal [4]. We propose to use the macrophages to deliver the nanoshells to areas of hypoxia within tumors where they will be irradiated killing the macrophage and the surrounding malignant epithelial cells.
Chemoprevention of Colon Carcinogenesis by Dietary and Medicinal Mushrooms
Principal Investigators: Jiri Adamec, Lead Scientist - Proteomics/Metabolomics Facilities,
Bindley Bioscience Center-Discovery Park, Purdue University
Daniel Sliva, Ph.D, Department of Medicine/Methodist Research Institute,
Indiana University Cancer Center
Abstract
The objective of our project is to evaluate chemopreventative effect of medicinal and edible mushroom extracts against dietary-and inflammation-induced colon carcinogenesis. Our central hypothesis is that the extracts from an old Asian medicinal mushroom, Ganoderma lucidum (GLT) and extract from a common edible Oyster mushroom Pleurotus ostreatus (OME) will inhibit colon carcinogenesis induced by food born mutagenic chemicals (identified in cooked meat and fish) and by inflammation. We hypothesize that GLT and/or OME specifically modulate inflammation-dependent activation NF-kappaB/AP-1 and beta-catenin signaling, thereby inhibiting the promotion and metastasis of colon carcinogenesis.
We have formulated this hypothesis on the basis of our preliminary data clearly demonstrating that G. lucidum triterpene fraction (GLT): (a) inhibits proliferation and invasiveness of colon cancer cells; (b) suppresses TNF-induced NF-kappaB activation; (c) is well tolerated and not toxic in mice. Furthermore, daily gavage of GLT to rats clearly demonstrates that the majority of compounds identified in GLT by microLC-ESI-MS, which we recently developed, are transported and retained in colonic mucosa. In addition, our studies also showed that an extract from Oyster mushroom (OME); (a) suppresses proliferation of colon cancer cells through cell cycle arrest at the G0/G1 phase, (ii) inhibits migration of colon cancer cells, and (iii) orally administered OME is not toxic for mice. Our rationale for these studies is that the development of scientifically based evidence to support the anti-inflammatory and anti-cancer effects of Ganoderma lucidum and Pleurotus ostreatus would provide a foundation for the future in vivo studies and clinical trials to test medicinal and dietary mushroom as a nutritional chemopreventive agent. We propose the following specific aims:
The primary impact of our anticipated findings on public health will be evidence-based scientific verification of the anti-cancer effect of mushroom extracts which may help to establish the dietary use of mushrooms to prevent colon cancer.
A Randomized Pilot of Breast Cancer Survivor Patient Activation Intervention
Principal Investigators: Cleveland Shields, Ph.D, Child Development and Family Studies, Purdue University
Victoria Champion, DNS, FNA, Associate Dean for Research,
Mary Margaret Walther Distinguished Professor of Nursing,
Director of Cancer Control,
Indiana University
Abstract
BACKGROUND: Breast cancer survivors face worries and fears about recurrence and long term side effects. These patients have been in intensive treatment for months, but as their active treatment ends they shift from weekly or even daily contact (when they were receiving radiation treatment) with physicians and nurses, to contact every 3 to 6 months. Patients' ability to make the most of these less frequent visits has an impact on their quality of life, symptoms of post traumatic stress, and satisfaction with medical care. Dr. Champion's pilot work has found that more distressed patients report having difficulty talking about these issues with their physicians. The literature shows that patients frequently bring up emotional distress through clues, making it difficult for physicians to identify and respond appropriately.
SPECIFIC AIMS: 1) To examine the effect of a worries and concern prompt sheet vs usual care on patient perception of the visit. 2) To examine the effect of a worries and concern prompt sheet on patient distress. 3) To examine the effect of a worries and concern prompt sheet on physician satisfaction with the visit.
METHOD: We propose to conduct a pilot randomized trial of a patient activation intervention designed help patients directly express their worries and emotional concerns about recurrence and long-term side effects of their cancer treatment.
PARTICIPANTS: Sixty patients will be randomly assigned to receive the patient activation intervention versus usual care. For this pilot, we will recruit patients who have participated in Dr. Champion's ACS study with whom we already have an ongoing relationship. Patients will receive a letter from their oncologist inviting them to participate or to opt out of further contact. A week later, a research assistant will contact the patient by phone to ascertain their interest in participating. Patients, who agree verbally to participate, will be sent consent documents to sign and return.
We will randomize patients to receive either usual care or a question prompt sheet 1 week prior to their medical visit. The research assistant will call the patient and coach them to complete the question prompt sheet and instructing them to take the completed prompt sheet to their upcoming visit. One week after the visit, all participating patients will complete a questionnaire over the phone with the research assistant.
MEASURES: We will assess patient perceptions of the visit, patient distress, and physician satisfaction.
SIGNIFICANCE: Our study builds on previous research in question prompt sheets and will provide important knowledge to inform interventions that increase quality of life in younger breast cancer survivors. The intervention we are developing and testing could be adopted as standard practice in oncology settings with cancer survivors. Helping survivors control their fears and anxieties will help them manage ongoing cancer related difficulties. This intervention could be easily adapted to a Web interface that could be accessed prior to a patient visit when Web access is universally available making the intervention extremely cost-effective and portable. Finally, this study focuses on a population of cancer survivors that have been underserved-those diagnosed at 45 and younger.
The mission of the Oncological Sciences Center is to seek new opportunites, forge new partnerships, and nurture new relationships to advance cancer research beyond the laboratory.
Oncological Sciences Center
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