Grant of the Month

2007

December
PIs: William Wagner, PhD
Title: Cardiopulmonary Organ Engineering
Description: Researchers are currently working to develop tissue constructs that could be utilized to repair congenital heart defects and to recover function lost to myocardial infarction. The general approach involves optimizing the combination of precursor cells with synthetic scaffolds and culturing methods to achieve the functionality required for the given application. The research team working on this project includes cardiothoracic surgeons, bioengineers, polymer chemists, cell biologists and electrophysiologists.
Source: NIH-Bioengineering Research Partnership
Term: Year 5 continuation
Amount: $1.18 million

November
PIs: Satdarshan P. Singh Monga, MD
Title: Beta-catenin in the growth of hepatocellular cancer
Description: Hepatocellular cancer (HCC) is a disease of poor prognosis. Identifying novel molecular aberrations might present opportunities to identify new therapeutic targets. Due to the similarities between the processes of development and cancer, we used early developing livers to identify genes that might play a primary role in HCC. Platelet-derived growth factor receptor-alpha (PDGFRalpha) was identified from microarray using early developing mouse livers. Expression of PDGFRalpha and its upstream effectors, PDGF-AA and PDGF-CC, were examined in HCC tissues (n = 43) by Western blot, real-time PCR, and immunohistochemistry. Finally, effect of anti-PDGFRalpha antibody (mAb 3G3, ImClone Systems, Inc.) was examined on human hepatoma cells. A high expression of PDGFRalpha was observed during early liver development. HCCs (17 of 21) revealed cytoplasmic PDGFRalpha and activated PDGFRalpha (phospho-Tyr(754)) by immunohistochemistry. Additional HCCs (14 of 22) showed elevated PDGFRalpha levels when compared with the adjacent normal livers by Western blots. Of these 14 patients, 3 showed increased PDGFRalpha gene expression, 3 showed elevated PDGF-AA, and 4 had higher PDGF-CC levels in the tumors compared with adjacent livers. Multiple hepatoma cell lines, when treated with mAb 3G3, showed significant decreases in cell proliferation and survival (P < 0.05). In conclusion, approximately 70% of HCC tissues had elevated PDGFRalpha levels due to diverse mechanisms. PDGFRalpha inhibition in hepatoma cells led to diminution of tumor cell survival and proliferation and thus might be of therapeutic significance.
Source: National Cancer Institute
Term: 3 years
Amount: $1.2 million

October
PIs: Michael S. Sacks, Ph.D.
Co-PIs: Dr. Joyce Bischoff, Dr. David Brown, Dr. Danielle Gottlieb, Dr. John Mayer, Dr. Robert Padera, Dr. Andrew Powell, Dr. Virna Sales, Dr. Frederick Schoen, Dr. George Stetten
Title: Mechanisms of In-Vivo Remodeling in Tissue Engineered Heart Valves
Description: Using autologous cells and biodegradable polymers, tissue engineered pulmonary valves (TEPV) have been fabricated and have functioned in the pulmonary circulation of growing lambs for up to 20 weeks, with tissue evolving into a differentiated layered structure resembling that of native valve.  More recent studies have demonstrated that use of bone marrow mesenchymal stem cells (BMSC) and PGH/PLLA scaffolds produce functioning implants for up to 8 months in growing lambs which also demonstrated in-vivo structural evolution.  These studies have demonstrated the feasibility of engineering pulmonary valve (PV) leaflets and segments of main pulmonary artery (PA) in-vitro.  Both stuctures have functioned well without thrombosis.  Moreover, both the gross and microscopic characteristics of the TEPV structures began to approximate those of normal tissues, strongly suggesting that cell phenotypes evolved in a directed fashion to remodel the valvular and vascular tissue.  The goal of the current research program is to quantify and simulate tissue remodeling events that occur post-implantation, and to understand the factors that influence the remodeling rate and the quality and architecture of the ultimate tissue.  Specifically, we hypothesize that TEPV implant remodeling is primarily mediated by the level of in-vivo mechanical stimuli to the interstitial cells and developing ECM.  Mechanical stimuli will affect the rate of scaffold degradation and the degree of post-implant cellular ingrowth.
Source: NIH-National Heart, Lung and Blood Institute
Term: 4 years
Amount: $3.3 million

September
PIs: Jean Latimer
Co-PIs: Stephen Grant and Michael Epperly
Title: Biomarkers of invasiveness using isogenic and progressive human ductal carcinoma in situ cell lines
Description: We have developed a novel tissue engineering system for Human Mammary Epithelial Cells (HMEC), both normal and malignant. This system allows for the long-term (>3 months) establishment of normal primary cultures that begin as 3-dimensional mammospheres. These mammospheres differentiate into complex branching ducts and lobules, i.e. reform the plumbing system of the breast in culture. The ability to form ductal structures from human breast tissue is absolutely unique to our laboratory. Tumor cells lack the ability to form these epithelial architectures. From these robust primary cultures we have generated 2 unprecedented DCIS cell lines from the same white patient¹s breast without the use of viruses or telomerase expression.
Recently we have also created another set of set of DCIS and contralateral cell lines from an Asian woman. The DCIS lines show chromosomal abnormalities consistent with a malignant phenotype (although not as abnormal as MCF7). Our initial goal was to use matched sets of DCIS cell lines to identify biomarkers consistent with early stages of breast oncogenesis by comparing the contralateral cell lines with DCIS cell lines.We have now developed a set of biomarkers from microarray analysis that correlate well with invasiveness potential in these isogenic sets of cell lines. Validation of these biomarkers has been obtained using additional stage I and II cell lines we have also established, with known clinical recurrence patterns in the patients.  In aim 2 we will validate the invasiveness of these cell lines by placing all DCIS and isogenically matched contralateral and non-tumor cell lines into transwell chambers and immunocompromised mice.  Subcultures of the most motile cells will be analyzed in each case with microarray to validate the markers most associated with a motile and invasive phenotype.  Concurrently, we will establish similar matched sets of DCIS cell lines from African American women. In women diagnosed with DCIS 1973-2000, 9.2% were White, 10.2% Black, 7.3% American Indian, 15.0% Asian/Pacific Islander, and 11.5% Other.   Black women had a relative risk of mortality of 1.35 compared to white women, while Asian women had a reduced relative risk of 0.74, American Indian women had a RR of 0.95. The results we have obtained on the white and Asian women will be compared with those of the AA women in our study to assess whether DCIS, is associated with a higher risk of invasiveness in AA women. DCIS is one of the most commonly diagnosed forms of breast cancer (BC) because it is visible on mammograms.  At the same time it is one of the least understood forms of BC with the likelihood of becoming invasive BC unknown.  We have identified  biomarkers associated with the early stages of breast oncogenesis using these unique progressive systems.  We will further develop a population specific set of invasiveness markers for DCIS to assist in the tailoring of treatments for DCIS in African American women.
Source: Komen for the Cure Agency
Term: 2 years, 5/1/07-4/31/09
Amount: $600,000

August
PIs: Kacey G. Marra, PhD
Co-PIs: Douglas Weber, PhD
Title: Surface Modified Polymer Conduits for Peripheral Nerve Repair
Description: The project seeks to increase the understanding of the effects of biomaterial surface properties on nerve regeneration, and also to enhance the integrated research and education activities of graduate, undergraduate and pre-college students.  The plan includes outreach programs and innovative research in the exciting areas of biomaterials and tissue engineering.  Specifically, the research plan is focused on developing biodegradable conduits with modified inner luminal surfaces that enhance peripheral nerve repair over long gaps, and the educational plan is focused on enhancing and maintaining the involvement of women and underrepresented minorities in engineering.
Source: NSF-Biomaterials Program-Division of Materials Research
Term: 3 years
Amount: $300,000

July
PI Stephen Badylak, DVM, PhD, MD
Co-PI(s)
  • Susan Braunhut, Ph.D., professor of biological sciences at the University of Massachusetts at Lowell
  • Lorraine Gudas, Ph.D., chairman of the pharmacology department and Revlon Pharmaceutical Professor of Pharmacology and Toxicology, Weill Medical College of Cornell University, New York City
  • Ellen Heber-Katz, Ph.D., professor, molecular and cellular oncogenesis program, The Wistar Institute in Philadelphia
  • Shannon Odelberg, Ph.D., assistant professor, departments of internal medicine and neurobiology and anatomy, University of Utah, Salt Lake City
  • Hans-Georg Simon, Ph.D., a developmental biologist and assistant professor of pediatrics, Children’s Memorial Research Center and Northwestern University in Chicago
Title “Mammalian Limb Restoration”
Summary The regenerative ability of adult human tissues, organs, and appendages is typically very limited.  The default mechanism of wound repair in humans and most other mammals is characterized by scar tissue formation.  However, there is evidence for some site-specific regeneration-like processes during mammalian embryologic development and during the early postnatal period.  In addition, there is lifelong self-renewal capability for selected cell populations such as hematopoietic cells, intestinal epithelium, and hepatocytes. In contrast, urodele amphibians possess extraordinary abilities to regenerate lost structures, such as the limbs and tail, throughout their lifetime.   These regenerative processes are dependent upon the formation of a blastema at the site of injury.  This regeneration blastema is comprised of a self-organizing pool of proliferating progenitor cells genetically programmed to develop into a phenocopy of the lost structure.  The blastema carries its own extracellular matrix and its own gene expression signature.  The work described in this project will attempt to unlock the regenerative potential in humans by determining the genetic signature of the developing blastema and attempting to recreate portions of the fetal development process in humans. The research will involve several milestones including identification of cells that participate in the formation of a blastema-like structure in mammals, the spatiotemporal location of such cells during the remodeling process and the identification of bioactive molecules that induce, maintain, and complete such a process.  The culmination of this work would eventually be the application of these identified mechanisms and events to the injured mammal in a mouse model. A highly interdisciplinary research team has been developed with expertise in developmental biology, molecular biology, matrix biology, pharmacology, immunology, and with training in medicine, veterinary medicine, physics, and computational methods of data mining.  Significant preliminary data has been generated to support the fundamental approach.  Well defined milestones have been identified and a management scheme has been established that assures close collaboration among the principal investigators and their respective laboratories at six different institutions.
Source DARPA (W911NF-06-1-0067); Year 2 funding ($3,605,738)
Term 6/15/07 – 6/14/08

June
PI Billy W. Day, Ph. D. and Jean J. Latimer, Ph.D.
Title “Quantitative proteomics of nuclear matrix proteins in novel human ductal carcinoma in situ model systems”
Summary Background: Ductal carcinoma in situ (DCIS) is the earliest identifiable breast cancer lesion. Because DCIS is a pre-invasive malignancy, a better understanding of if and how it may progress to invasive disease will allow determination of which patients to treat aggressively and avoid unnecessary aggressive procedures. Once determined, these differentially expressed proteins may be used as biomarkers or therapeutic targets, as well as help determine the paths by which normal cells progress to DCIS plus provide a better understanding of breast carcinogenesis and ways to prevent it. Tumor grade, size and presence of necrosis are currently used to make clinical decisions regarding DCIS.

These have not proven to be good predictors, however, as low-grade DCIS often progresses to invasive disease. Proteins that hold promise for a better understanding of DCIS are those in the nuclear matrix (NMPs). The nucleus is a cellular landmark in the pathology of cancer. NMPs in part help to determine the nuclear shape and processes, have been identified as informative markers of disease states in a variety of cancers, and their detection in the serum and urine supports investigation of them in DCIS. NMPs have been investigated in invasive breast cancer, and several have been identified as unique to malignant specimens. Investigation of the NMPs in DCIS is therefore warranted. Prof. Latimer has over the past decade derived several unprecedented DCIS- and breast reduction mammoplasty-derived cell lines, generated without the use of transforming agents from clinically well-defined patients. In combination with the Day lab's modern proteomics technologies, we will thoroughly investigate DCIS. The DCIS samples, both invasive and non-invasive, gave rise to cell lines from tumors and nontumor adjacent tissue, and Prof. Latimer also has cell lines derived from contralateral non-diseased breast samples. All of the lines are characterized by karyotype, array-based comparative genomic hybridization, growth rates, breast epithelial markers, and functional DNA repair capacity.
Hypothesis: Detectable differences exist between invasive and noninvasive DCIS at the protein level, particularly in the nucleus, and these differences are distinct from those demonstrated at the nucleic acid level.

Specific Aims: In this 2-year study, we will characterize these lines by examining their differential protein expression. In Aim 1, we will grow cultures of the various cells in quantities necessary for replete proteomics analyses, isolate NMPs from each, then employ two of our several proteomic techniques, namely difference two dimensional gel electrophoresis (DIGE) and isobaric tags for relative and absolute protein quantitation (iTRAQ), each followed by mass spectrometric protein identification. These two methods offer orthogonal means of quantitation to help verify results. In Aim 2, the results will be further evaluated with Western blotting and RNAi.
Study Design: By comparing the DCIS sample with the pathologically normal adjacent tissue, in addition to the contralateral normal breast sample, we will be able to determine markers specifically expressed by DCIS and investigate their role in the disease, including invasiveness.

The mammoplasty cultures will be used as truly normal comparative controls.
Impact: If we understand the changes in the cells that give rise to DCIS, we may be able to identify the origins of breast cancer and develop preventative agents.

Source DOD Synergy Award (only 10 were given) $800,000
Term 2-year study

May
PI Stephen F. Badylak, D.V.M., M.D., Ph.D.
Title Biomechanical Soldier Treatment and Regeneration Consortium (STRaC):  Preclinical Study to evaluate the use of UBM-ECM for Partial Thickness Skin Wounds
Summary This award provides support to evaluate the use of a biologic scaffold harvested from the porcine urinary bladder (UBM) for the treatment of burn injuries.  The clinical trial will be conducted at the Institute for Surgical Research at Fort Sam Houston in San Antonio, TX.  The pilot study will involve the treatment of donor sites for patients that require split-thickness skin grafts.  The subsequent trial will involve treatment of the primary wound itself, thus obviating the need for a donor tissue harvest.
Source PTEI via DOD (STRaC)
Term 10/01/06 – 09/30/07

April
PI Michael Sacks, PhD
Co-PI(s) William Wagner, PhD
Title Biomechanical Optimization of Tissue Engineered Heart Valves
Summary The focus of this competitive renewal grant is a comprehensive biomechanical evaluation of the in-vitro phase of engineered tissue heart valve development.
Source NIH/NHLBI: R01 HL68816-01
Term 2/01/07– 12/31/11

March
PI Derek C. Angus, M.D., M.P.H
Co-PI(s) Donald M. Yealy, M.D and Mitchell P. Fink, M.D.
Title Protocolized Care for Early Septic Shock (ProCESS)
Summary The Protocolized Care for Early Septic Shock (ProCESS) study will attempt to determine if there is a "golden hour" in the management of sepsis and septic shock when a prompt, rigorous, standardized treatment regimen can be used to improve clinical outcomes and halt the cascade of events that often lead to organ failure and death. The study takes a cue from the realm of coronary care, which has significantly reduced mortality from acute coronary diseases and dramatically reduced the costs of care by determining such best practices.

The project is designed to generate comprehensive data on the clinical and biological aspects of standardized treatment for septic shock – data that can have an immediate impact on and improve the care of the critically ill. The trial, to be conducted at several leading hospitals around the country, will enroll up to 2,000 participants who present to the emergency department with septic shock. Participants will be randomized to receive alternative treatment protocols involving intravenous fluids, drugs that reverse the shock and hemodynamic monitoring during the first six hours of care.

The protocols will be evaluated on three measures: clinical effectiveness as evidenced by improved mortality rates; effectiveness in reducing concentrations of biological markers that are associated with the four fundamental pathways of sepsis-related organ dysfunction – cellular hypoxia, oxidative stress, inflammation and coagulation/thrombosis; and cost effectiveness.

Sepsis is among the top causes of death in the United States, affecting 750,000 Americans each year, of which 30 percent die. It also is one of the most expensive diseases, with a cost to U.S. hospitals of $17 billion each year.
Source NIH/National Institute of General Medical Sciences

February
PI See below
Title National Tissue Engineering Center (Multiple Awards)
Summary Regenerative Medicine Approach to the Treatment of Abdominal Compartment Syndrome in a Dog Model
PI: Stephen F. Badylak, MD, DVM, PhD
Abdominal compartment syndrome (open abdomen) is occurring with increasing incidence in wounded soldiers requiring in-theater “damage-control laparotomy”.  The inability to close the fascia of the abdominal compartment following surgery results in prolonged open abdomen in these patients.  Current methods of closure involve synthetic materials, high rates of infection, high morbidity, and a poor outcome.  The present proposal will evaluate a regenerative medicine approach in which the abdominal wall will be reconstructed with functional musculotendinous tissue by the use of an inductive bioscaffold composed of porcine derived extracellular matrix (ECM).  The study will be conducted in a dog model that mimics the human clinical situation.

Signaling and Cellular Strategies of Injectable Biomimetic Matrices for Craniofacial Bone Tissue Engineering
PIs: Charles Sfeir, DDS, PhD and Prashant Kumta, PhD
This project addresses materials development primarily involving the synthesis and characterization of novel injectable bone cement composites containing nanostructured carriers of signaling molecules and protein, as well as an investigation of the signaling properties of specific mineralized tissue extracellular matrix proteins (ECM) and the isolation and purification of stem cells.  These ECM and stem cells will be incorporated into the newly synthesized injectable bone cement materials to form a novel smart biomimetic matrix, the combination of which will be assessed in an animal model to investigate their potential for craniofacial bone tissue engineering. 

Rapid Engineered Autologous Blood Vessels
PI: William Wagner, PhD and David Vorp, PhD
Currently available grafts for small diameter vascular replacement or bypass are fraught with limitations, especially for soldier care purposes.  It is clear that new alternatives are needed, and it is widely felt that the burgeoning field of tissue engineering will be crucial to the development of these new vascular grafts.  Most vascular tissue engineering approaches rely on some sort of scaffold where cells are incorporated. However, many aspects remain unclear regarding the biomaterial properties of the scaffolds, and the cell source used to populate them. A novel poly (ester urethane) urea elastomeric scaffold has been developed and shown to have great potential for cardiovascular tissue engineering applications. Because of their multipotentiality and availability as an autologous cell source, progenitor cells are considered as the ideal source for tissue engineering. Our preliminary data, where a rapid incorporation of progenitor cells within the bioerodible polymer is achieved through a novel seeding technique, shows that a construct fully-seeded with viable cells can be achieved in a very short period of time.

Accordingly, the goal of this project is to develop a novel, bioresorbable scaffold, bulk-seeded with human stem cells, and cultured acutely in-vitro so that an optimal implant is available in a short amount of time.  To this end, we propose two specific aims for this one-year project:

  • Develop a rapidly fabricated, bioerodable polymer-based vascular graft bulk-seeded with human stem cells, and
  • Test the mechanical properties, structure and functionality of the vascular graft from Specific Aim 1following an acute, optimized culture period to determine its readiness for in-vivo implantation for further development into a bio-equivalent vascular substitute.
Source National Tissue Engineering Center

January
PI John A. Kellum, MD
Title Biological Markers of Recovery for the Kidney (BioMaRK)
Summary Investigate the role of inflammation, as well as other factors in recovery from acute renal failure (ARF). This project, called Biological Markers of Recovery for the Kidney, or BioMaRK, will examine how such factors influence survival as well as recovery of kidney function. The study will assess how certain inflammation markers relate to clinical outcomes and build a risk-prediction model based on clinical variables and those biomarkers. The results of this study could potentially lay the foundation for the development of ARF treatment therapies, particularly those designed to enhance organ recovery.
Source NIH- National Institute of Diabetes and Digestive and Kidney Diseases
Term 5 Years

2008 | 2007 | 2006