Radiation Oncology

1. Senior Research Staff

Steve Brown, Ph.D.
Svend Freytag, Ph.D.
Jae Hoe Kim, M.D.
Benjamin Movsas, M.D., Chair

2. Research Summaries

Principal Investigator: Brown, Steve, Ph.D.
Core C: Training and Education of Center Grant: Post-Irradiation Intervention to Mitigate and Treat Non-Hematological Injuries (NIH U19AI1067734 Subcontract)

The Training and Education Core will focus on the post-doctoral trainees who will be involved in the Research Projects and to a lesser extent on the faculty researchers whose training is not in radiation biology. There will be both a laboratory and a didactic component. The laboratory training will consist of hands-on research under the direction of Project Leaders. Trainees will gain experience in the use of assays, methods, reagents, animal models, and technologies to study the mitigation and/or treatment of radiation injuries. Included in the Core is provision for trainees to spend some of their time in labs of more than one Project. It is also expected that the trainees will prepare the results of their studies for peer-reviewed publication. The didactic program will cover the principles governing the response of normal tissue to radiation through a series of talks from experts from within and outside the Center. The lecture series will consist of a videotaped seminar series along with a self-test module. In addition, trainees will be expected to attend laboratory journal club departmental/institutional speakers programs and ongoing radiation biology lectures being delivered to radiation oncology residents in order that the trainee gains a broader perspective of his research and areas of specialties. A website based at Henry Ford Health Systems, but shared between members of the consortium, is a fundamental part of the Training and Education Program. In summary, the purpose of this core is to train the next generation of normal tissue radiobiologists.

Principal Investigator: Brown, Steve, Ph.D.
PPG Project Core C: The Tumor/Cell Biology and Histology Core (NIH P01CA09701201A) The Tumor/Cell Biology and Histology Core (Core C) will have the following functions:

1. maintain inventory of cell lines,
2. expand tumor cells for implantation into animal subjects,
3. perform necropsies, routine histology and immunohistochemistry on tissues,
4. prepare spleenocytes and hepatocytes for in vitro cytolytic CTL assays,
5. analyze prostate biopsies for therapeutic gene expression, and
6. monitor neutralizing antibodies to adenovirus in patient's serum.

Principal Investigator: Brown, Steve, Ph.D.
PPG Project 2: Improved Gene Delivery and In Vivo Imaging (NIH P01CA09701201A)

Gene therapy is currently limited by the lack of a non-invasive and clinically useful means to monitor vector persistence and biodistribution and transgene expression in vivo. The goal of the current project is to test strategies devised to improve the spatial distribution of gene expression following the local administration of adenovirus-mediated gene therapy so that the entire organ receives a viral dose sufficient for clinical efficacy with as little virus as possible (to limit toxicity). A new reporter gene based on the sodium iodide symporter (NIS) will be characterized with regard to its ability to monitor therapeutic gene expression and contrasted with other measurements of catalytic activity under development. There are three specific aims of the proposed studies.

1. Compare the distribution of NIS reporter gene expression to that of CD/HSV-1 TK therapeutic gene expression. If it can be shown that NIS activity is a surrogate marker of therapeutic gene expression, NIS imaging could eliminate the biopsy currently used to assess transgene expression and allow for dynamic and whole body monitoring of transgene expression.
2. Evaluate new vector formulations and injections conditions designed to optimize placement of the adenovirus within the prostate to obtain maximal organ coverage. This work will provide the scientific basis for a clinical trial described in Project 3.
3. Contrast the prodrug-labeled approach of monitoring transgene expression to the NIS approach. This aim tests the hypothesis that the bystander effect can be measured and exceeds the area of gene expression. Bystander effect will be measured using autoradiography of bound radiolabeled prodrugs ([3H]-ganciclovir-MP, [14C]-5-FdUMP) and compared to iodide localization following transduction of NIS.

Principal Investigator: Freytag, Svend, Ph.D.
PPG Project Core A: The Administrative Core (NIH P01CA09701201A)

The Administrative Core (Core A) will provide the administrative support and oversight functions of the Program Project. The Administrative Core will consist of an Executive Committee comprised of the project and core leaders and a full time Grants Administrator. The Administrative Core will have the following functions and responsibilities:

1. review productivity,
2. review scientific directions,
3. foster collaborations and coordinate efforts among the projects and cores,
4. discuss problems and develop strategies for overcoming them,
5. monitor and manage fiscal aspects of the Program Project,
6. monitor and manage regulatory aspects of the Program Project,
7. coordinate interactions and administer correspondence with internal committees and external agencies, and
8. organize and support a seminar series in gene therapy.

Principal Investigator: Freytag, Svend, Ph.D.
PPG Project Core B: The Molecular Biology and Vector Core (NIH P01CA09701201A)

The Molecular Biology and Vector Core (Core B) will have the following functions:

1. maintain inventory of recombinant DNA vectors,
2. perform large-scale plasmid preps,
3. perform plasmid cloning,
4. perform HEK 293 cell transfections,
5. perform large-scale adenovirus preps,
6. characterize recombinant adenoviruses (laboratory and GMP grade),
7. analyze potency of adenovirus following each patient injection,
8. monitor presence of adenoviral DNA in patient's blood using PCR/Southern blot assay.

Principal Investigator: Freytag, Svend, Ph.D.
Molecular Gene and Radiation Therapies for Cancer (NIH P01CA09701201A)

The NCI-sponsored Program Project entitled "Molecular Gene and Radiation Therapies for Cancer" builds on the past preclinical and clinical accomplishments of the Department of Radiation Oncology's Gene Therapy Program. Their program has developed a novel gene therapy approach designed to improve the effectiveness of radiation therapy. They recently sponsored and completed two successful prostate cancer clinical trials at the HFHS that were a direct result of their research efforts.

The newly awarded Program Project is comprised of three projects and four cores that function as a highly integrated and comprehensive unit that will advance gene therapy technology on three fronts: 1) by developing better gene therapy products (Project 1), 2) by developing better means of product delivery and monitoring (Project 2), and 3) by evaluating the merit of these preclinical advancements in the clinic (Project 3). An important aspect of the two preclinical projects (Projects 1 & 2) is that all of the studies were designed to be translatable into the clinic. Project 3 describes three Phase I/II clinical trials that will examine the safety and efficacy of their novel gene therapy approach in combination with radiation therapy in patients with newly diagnosed prostate cancer using a "new and improved" gene therapy product.

Principal Investigator: Freytag, Svend, Ph.D. PPG Project
1: Second Generation Adenoviral Vectors for Cancer Therapy (NIH P01CA09701201A)

Our research program has developed a novel, trimodal gene therapy-based approach for the treatment of cancer. Our approach utilizes a cytolytic, replication-competent adenovirus (Ad5-CD/TKrep) to selectively and efficiently deliver a pair of therapeutic "suicide" genes to tumors. Our preclinical studies have demonstrated that the Ad5-CD/TKrep virus itself, via its cytolytic activity, has potent anti-tumor activity. The efficacy of Ad5-CD/TKrep viral therapy can be enhanced significantly by invoking two suicide gene systems (CD/5-FC and HSV-1 TK/GCV), which render malignant cells sensitive to specific pharmacological agents and importantly, sensitizes them to radiation. A major objective of Project 1 is to develop second-generation adenoviruses that may be more efficacious and less toxic than the parental Ad5-CD/TKrep virus. We will determine whether second-generation adenoviruses containing various E3 genes demonstrate greater anti-tumor activity in an immune-competent host relative to the parental Ad5-CD/TKrep virus. We will test the hypothesis that suppression of the host immune response by E3 genes will result in longer-term therapeutic gene expression and improved tumor control. Anti-tumor activity will be correlated with the duration of therapeutic gene expression in vivo and the extent, and nature, of the immune response. We will determine whether second-generation adenoviruses expressing a more catalytically active yeast CD/mutant HSV-1 TK(SR39) transgene results in better tumor control than the parental Ad5-CD/TKrep virus. We will examine the toxicity of second-generation adenoviruses in the immune-competent mouse following intraprostatic and intravenous administration. Finally, we will develop a series of second-generation adenoviruses expressing the human sodium iodide symporter (hNIS). We will test the hypothesis that expression of hNIS will enable virus-infected cells to take up (99m)TcO4- and (123)I and allowing for non-invasive monitoring of vector biodistribution and therapeutic gene expression in vivo. All of the second-generation adenoviruses developed in Project 1 will be used in the other projects of this Program.

Principal Investigator: Kim, Jae Hoe, M.D.
Mitigating/Treating Radiation-Induced CNX Injury with ACE Inhibitors and Statins (NIH U19AI1067734 Subcontract)

The central nervous system (CNS) is the most critical of all normal tissues for maintaining coordinated normal function. Following radiation exposure, the response of the CNS has been attributable to parenchymal and vascular damages including oligodendrocytes, neural progenitors, and endothelial cells. A dynamic process of radiation-induced death of target cells and subsequent secondary reactive inflammatory process is believed to lead to cell loss, tissue damage and functional deficits. Arguably of greater consequence than the physical damage is the psychological impact of CNS injury which could result in the loss of sight, paralysis or reduced cognitive function. The only investigational demonstration of pharmacological mitigation of CNS radiation damage was recently made by our group using the Angiotensin Converting Enzyme (ACE) inhibitor, ramipril (an FDA-approved drug). Of note, a reduction of injury assessed functionally and histopathologically was observed even when ramipril was administered weeks after the radiation exposure.

This project is based on the hypothesis that the suppression of oxidative stress resulting from multi-cellular interactions through a network of pro-inflammatory mediators would mitigate radiation-induced brain injury by specific pharmacological treatment. Three classes of drugs are well known to suppress oxidative stress and two of them, ACE inhibitors and their receptor blockers and statins are widely used in various cardiovascular disorders in humans. The project aims to address specific therapeutic roles of three classes of drugs, ACE inhibitors and receptor blockers, statins, and SOD mimetics, in all phases of the brain injury, i.e., acute, early delayed, and late delayed reactions. The primary goal is to demonstrate their efficacy and to optimize the use of drug dosage and timing of the administration to mitigate and treat the radiation brain injury. End points for evaluation are cognitive functions, visual function, MRI for permeability of the blood brain barrier, and histopathological and proliferative changes for neurogenesis, vascular and glial cells, using single doses of whole brain radiation of the adult rats. The positive findings from the proposed study will be readily translatable in humans, since both ACE inhibitors and statins are widely used in a variety of cardiovascular disorders in the clinics.

In summary, experimental studies suggest radiation-induced CNS injury can be treated. The goal of this project is to bring one or more of these experimental approaches into clinical practice.

Principal Investigator: Kim, Jae Hoe, M.D.
PPG Project 3: Suicide Gene and Radiation Therapy Clinical Trials (NIH P01CA09701201A)

The objective of Project 3 is to genetically manipulate cancer cells to increase intrinsic radiation sensitivity preferentially to tumor tissue with the ultimate goal of improving the outcome of radiation therapy. To achieve this goal, we will conduct a series of Phase I and II clinical trials that will test the general hypothesis that combining replication-competent adenovirus-mediated double suicide gene therapy with conformal radiotherapy can be applied safely in humans and will demonstrate superior efficacy compared to conformal radiotherapy alone. Specific Aim 1 describes a Phase I/II trial to determine whether replication-competent adenovirus-mediated double suicide gene therapy in combination with conventional dose (72 Gy) intensity modulated radiotherapy (IMRT) is superior to IMRT alone in patients with newly diagnosed, intermediate-to-high risk prostate cancer. The best second-generation adenovirus developed in Project 1 will be used. The primary endpoint will be local tumor control as determined by prostate biopsy status at two years. Other endpoints will be acute and late toxicity, early tumor control at six months and one year, and freedom from biochemical or clinical failure. Specific Aim 2 describes a Phase I/II trial to determine the safety and efficacy of replication-competent adenovirus-mediated double suicide gene therapy in combination with salvage IMRT in patients with locally recurrent prostate cancer. The best second-generation adenovirus developed in Project 1 will be used. Three cohorts of three to six patients will receive a single intraprostatic injection of adenovirus (10[12] vp) along with three weeks of 5-FC + vGCV prodrug therapy and an escalating dose (20, 26, 30 Gy) of IMRT. If there are no toxicity concerns at six months, a Phase II trial will be conducted in which patients will receive a single intraprostatic injection of adenovirus (10[12] vp) along with three weeks of 5-FC + vGCV prodrug therapy and the maximum tolerated dose (MTD) of IMRT. The primary endpoint will be local tumor control as determined by prostate biopsy status at two years. Other endpoints will be acute and late toxicity, and freedom from biochemical or clinical failure. Specific Aim 3 describes a Phase I trial that will evaluate the efficiency of therapeutic gene transfer in vivo using an improved vector formulation designed to enhance gene delivery. The efficiency of gene transfer and distribution of vector will be evaluated in patients with localized prostate cancer who are scheduled to undergo prostatectomy. These studies will generate important new knowledge that will provide the scientific basis for which future large-scale human trials will be based and may ultimately lead to better cancer treatments.

Principal Investigator: Lu, Mei, Ph.D.
PPG Project Core D: Biostatistics and Data Management Core (NIH P01CA09701201A)

The general function of the Biostatistics and Data Management Core (Core D) is to support the basic and clinical activities of the Program Project so that research studies are most efficiently designed, conducted, monitored, and analyzed. This includes data management, statistical modeling, formalization of hypotheses to be tested to ensure that valid conclusions can be drawn, design of clinical trials, and data analysis. All of these activities are carried out in collaboration with the laboratory and clinical investigators in all projects. The specific objectives are:

1. To consult and collaborate with investigators in the development of better adenoviral vectors and therapeutic gene systems;
2. To consult and collaborate with investigators on the development of imaging models to quantify the volume and intensity of gene expression in tissues and for optimizing therapeutic gene delivery;
3. To provide statistical consultation and collaboration in the planning, conducting, analyzing and reporting of Phase I/II clinical trials;
4. To coordinate project conduct and be responsible for data management and quality of clinical trials.

1. Senior Research Staff

Arbab S. Ali, Ph.D.
Manuel Brown, M.D., Chair
Michael Flynn, Ph.D.
Hamid Soltanian-Zadeh, Ph.D.

Principal Investigator Ali, Syed Arbah, Ph.D.
Differentiation of Glioma from Radiation Injury Using Cellular MRI

Despite extensive treatment strategies and investigations, the prognosis of glioblastoma multiforme is still poor. The main reason is the inability to delineate the margin of the tumor during routine investigations or during surgery. Moreover, current imaging modalities fail to differentiate, conclusively, the recurrent or left over tumor from radiation necrosis or necrotic tissues. For proper management and follow up it is utmost important to detect recurrent tumor as early as possible. Recently dendritic cell based vaccination and cytotoxic T-lymphocytes (CTL) are being considered for the treatment of recurrent glioma. In the animal models as well as in the early phases of clinical trials, CTL has been shown to accumulate in the glioma. By tracking the migration and homing of CTL it may be possible to differentiate recurrent glioma from radiation necrosis. Recently, using two FDA-approved agents, we formed ferumoxides-protamine sulfate complex and labeled any kind of mammalian cells. To examine whether labeled cells can be used as cellular probes to detect and differentiate physiological and/or pathological conditions, we have selected glioma and radiation injury models. It is hypothesized that in vivo magnetic resonance imaging (MRI) tracking of magnetically labeled CTLs will enable us to identify different patterns of accumulation and incorporation of labeled injected CTLs, thus allowing for differentiation between recurrent glioma and radiation injury. The goals of this study will be achieved by making glioma as well as radiation injury models in tumor bearing or control nude rats. Nude rats will be used to implant human U-251 glioma cell lines. In these rats we will test whether CTLs produced in vitro by U-251 cell lysate-pulsed dendritic cells can recognize the implanted human glioma and differentiate it from radiation injury. CTLs produced in vitro will be magnetically labeled using feridex- protamine sulfate complexes and the labeled cells will be injected into tail vein of the rats at different stages of their disease processes. These labeled cells, once incorporated into the tumors or areas of injury, can be detected as low signal intensity areas on in vivo and ex vivo MRI because of the susceptibility effects of iron oxides inside the cells. Serial MRI of tumors and radiation injured areas after injecting labeled cells at different time points will be obtained by a 7 tesla MRI system. The findings of MRI will be correlated with histology, and immuonohistochemical detection of CTLs. The results will also be compared among the animals of all groups. Early detection of recurrent or metastatic glioma as well as early differentiation of glioma from radiation necrosis will help clinician to tackle this devastating neurological malignant tumor. Early detection and differentiation of recurrent glioma from radiation injury/necrosis by noninvasive imaging technique is essential for the proper management of this devastating malignant disease. If these studies are successfully completed, the results can easily be translated into a clinical trial, where patients' own dendritic cells (differentiated from autologous monocytes and/or hematopoietic stem cells separated from peripheral blood mononuclear cells [PBMC]), can be used to produce cytotoxic T-lymphocytes (CTL) to target tumor cells for detection and differentiation of tumor from radiation injury/necrosis. Genetically engineered CTL can also be used. This magnetic labeling technique will also help investigators to track the injected CTL in the body as well as in the targeted areas by magnetic resonance imaging (MRI).

Principal Investigator Ali, Syed Arbah, Ph.D.
Cellular MRI in Glioma and Radiation Necrosis

Recently, using two FDA-approved agents, we formed ferumoxides-protamine sulfate complex and labeled any kind of mammalian cells. To examine whether labeled cells can be used as probes to detect and differentiate physiological and/or pathological conditions, we have selected glioma and radiation necrosis models. It is hypothesized that in vivo MR tracking of magnetically labeled cells will enable us to identify different patterns of accumulation and incorporation of labeled injected cells, thus allowing for differentiation between recurrent glioma and radiation necrosis. Glioma is a central nervous system neoplasm that typically shows hypervascularity. Unlike the surrounding normal regions of cerebral vasculature, areas of hypervascularity are typically permeable to contrast agents, and can thus be detected by contrast-enhanced MRI or CT. However, areas of radiation necrosis can also show enhancement due to active inflammatory reactions and increasing vascular permeability. Thus, distinguishing recurrent glioma from radiation necrosis becomes problematic if only changes in vascular permeability and/or volume are considered. One distinguishing characteristic, however, is that there is little active angiogenesis at the site of radiation necrosis. By determining the differential migration and incorporation patterns of labeled endothelial progenitor cells (EPCs) at the site of glioma, it should be possible to differentiate between radiation necrosis and recurrent glioma. If this proves feasible, a translation into clinical trials can quickly follow, employing autologous labeled EPCs. These labeled cells, once incorporated into the tumors or areas of necrosis, can be detected as low signal intensity areas on in vivo and ex vivo MRI because of the susceptibility effects of iron oxides inside the cells. These objectives will be achieved by obtaining serial MRI of tumors and radiation necrotic areas after injecting labeled cells at different time points. The findings on MRI will be correlated with histology and different markers of endothelial cells. Angiogenic factors will also be assessed by immunohistochemistry at the site of accumulated EPCs in tumors or radaition necrosis. Early detection of recurrent or metastatic glioma as well as early differentiation of glioma from radiation necrosis will help clinician to tackle the devastating neurological disease.

Principal Investigator: Soltanian-Zadeh, Hamid, Ph.D.
Integrated Image Analysis System for Epilepsy (NIH R01 EB00245001A1)

With the ever-increasing role of medical images in diagnosis, treatment, and evaluation of treatment effects, extraction of quantitative information from these images and efficient use of the results have become a necessity. In recent years, others and we have developed novel two-dimensional (2D) and three-dimensional (3D) deformable models for a variety of image analysis applications in medicine and industry. We have also developed reliable automated methods for defining the initial shape of the model for segmentation and characterization of hippocampus from magnetic resonance imaging (MRI). These methods need to be extended and feature extraction methods developed to segment and characterize (i.e., determine multi-parametric intensity distribution, texture, shape, surface area, and volume of) brain structures such as hippocampus, amygdala, red nucleus, substantia nigra, globus pallidus, putamen, corpus callosum, and thalamus from MRI. In addition, new databases need to be developed to hold the results with other clinical information (e.g., textual data) in a manner that can be searched, retrieved, and queried conveniently from any computer station. The goal of this project is to develop novel approaches for the above needs. Developments will be done in the context of an important biomedical application and will localize, segment, and characterize hippocampus from MRI. The proposed database will be able to evaluate correlation between a variety of risk factors and post-operative outcomes. The methods will be tested; evaluated, and validated, using simulated images and clinical studies of epileptic patients. Clinical diagnosis based on EEG studies and surgery outcome will be used as "gold standards" for evaluation and validation of the image analysis methods. The proposed research will be a breakthrough in the development and application of computerized methods for medical image quantitation and object characterization, and will advance image analysis science in the direction of integrating knowledge-based systems, deformable models, texture analysis, and database technology. The proposed approach is applicable to the identification, segmentation, and characterization of other biological structures (e.g., lung, liver, kidneys, cells, neurons). It is also applicable to virtually any image analysis task for which object quantitation and characterization are used.

The National Institute of Neurological Disorders and Stroke (NINDS) is seeking to develop a Neurological Emergencies Treatment Trials (NETT) Network of Clinical Site Hubs. The Hubs will work with the NETT Clinical Coordinating Center to improve outcomes for patients with neurological emergencies through research. The clinical site Hubs will be regional consortia of emergency departments (ED) that will recruit patients and carry out phase III clinical trials. The Henry Ford Health System (HFHS) has a long commitment to both basic science and clinical research in neurological disorders and is a NINDS designated Stroke Center with 18 years of continuous funding. This proposal is a natural extension of our current work. The objective of this proposal is to demonstrate that the HFHS is an ideal Hub for the NETT Network. The specific aims are to create a flexible Hub with seven "spokes" at HFHS, to participate in research that improves patient care, to help create an enduring research network, to allow multi-specialty collaboration, and to foster research skills in young investigators that will allow them to pursue a career in clinical research. The Hub will be designed around the HFHS because it is a regional, integrated system of 6 area hospitals, 9 emergency departments, including 4 JCAHO certified stroke centers, and 36 clinics. It also includes a closed medical group of over 800 physicians with over 2.5 million outpatient visits and over 350,000 emergency department visits annually. The HFHS serves a wide variety of minorities, women, and children. The system is integrated through a central IRB, shared electronic medical record, standardized patient care and referral protocols, communication systems, and an integrated governance and leadership structure. A dedicated ambulance service (Superior Ambulance) interconnects the system. This proposal will use three hospitals and seven EDs. The method of implementation will be the Ford Neurological Emergencies Cross- disciplinary Team (Ford NEXT). This 28 member multi-specialty team with expertise in Emergency Medicine, Prehospital Care, the Neurosciences, Neuro- Intensive Care, Neuro-interventional Radiology, Trauma, Pediatrics, and Rehabilitation will implement and completely manage the clinical trials. The importance of this proposal is that the HFHS hub will build on a well established neuroscience foundation, rapidly complete studies in neurological emergencies, and improve patient care and outcomes.

This is a multidisciplinary consortium joining experts from the fields of clinical medicine, proteomics, mass spectrometry and biodiagnostics seeking to discover promising biomarkers for the identification of patients with severe sepsis and severe CAP.  The goals are to design a one timepoint assay to be developed into a point of care assay using an immunodiagnostic platform.  A combination of proteomic techniques with clinical data to study patients with severe sepsis and those at risk for a complicated course of CAP will be utilized.  By identifying patients in earlier stages of sepsis, interventions may be instituted to prevent progression of septic shock and multiple organ failure.