Title: Integrated Clilnical and Research Systems for Diabetic Foot Wound Care
Description: Diabetes is a common, complex, and costly disease affecting 9.4% (30.3 millions) of Americans. It remains the 7th leading cause of death in the United States, contributing to over 250,000 deaths annually. Diabetic foot ulcers (DFU) are the most frequently recognized complication in diabetics with an incidence of 6% in the diabetic global population, 6% among Medicare diabetic beneficiaries, 5% among diabetic U.S. veterans and a lifetime incidence of foot ulcers between 19% and 34% in diabetics. The natural history of a diabetes-related foot ulcer is devastating. More than half of ulcers become infected and approximately 20% of moderate or severe diabetic foot infections lead to amputation. Mortality after diabetes-related amputations is greater than 70% at 5 years for all patients with diabetes, which is 2.5 times higher than in diabetic patients without a foot ulcer. This proposal is designed to establish a clinical research unit (CRU) at the University of Pittsburgh Medical Center that integrates high quality care delivery seamlessly with outstanding clinical research. The CRU will then be a participating site in the NIH consortium studying biomarkers for diabetic foot ulcer healing. Our central hypothesis is that we can address the major challenges of diabetic foot ulcer clinical research through the seamless integration of wound center clinical operations with research operations. Our specific aims are: Aim 1: Establish a unified recruiting and retention system integrated with the clinical operations of our wound care service line, and linked to our EPIC scheduling system and electronic medical record (EMR). Aim 2: Establish integrated research quality systems, linked to highly standardized clinical pathways, and supported by a wound Informatics and Data Core connecting our 8 wound care centers.
Description: Drs. Glorioso and Cohen will share a two-year ACGT grant to support a study to develop a cancer vaccine for melanoma. Their research builds on previously successful results using a tumor-targeted, actively replicating herpes virus to infiltrate cancers and stimulate an immune system assault. Dr. Glorioso calls the methodology a “heat-seeking missile that targets metastatic cancer for destruction.” The treatment doesn’t stop there. Once the cancer is eliminated, the vaccine inserts an immunity barrier to protect against recurrence. Melanoma is among the most deadly cancers and this treatment offers new hope.
Title: Three-dimensional organoid models to study breast cancer progression
Description: Approximately 20% of breast cancers detected through mammography are pre-invasive Ductal Carcinoma in situ (DCIS). If left untreated, approximately 20-50% of DCIS will progress to more deadly Invasive Ductal Carcinoma (IDC). No prognostic biomarkers can reliably predict the risk of progression from DCIS to IDC. Similar genomic profiles of matched pre-invasive DCIS and IDC suggests that the progression is not driven by genetic aberrations in DCIS cells, but microenvironmental factors, such as hypoxia and metabolic stress prevalent in DCIS, may drive the transition. We need innovative models to investigate how to halt steps of DCIS progression to invasive phenotypes and subsequent metastasis from the primary site. This proposal directly addresses this unmet need by developing a novel three-dimensional in vitro organoid model that recapitulates key hallmarks of DCIS to IDC progression: tumor-size induced hypoxia and metabolic stress, tumor heterogeneity and spontaneous emergence of migratory phenotype in the same parent cells without any additional stimulus. A tangible advantage of the proposed organoid models is the ability to precisely and reproducibly study how the hypoxic microenvironment induces tumor migration in real time and in isolation from non-tumor cells present in vivo, providing unique opportunity to define tumor-intrinsic mechanisms of DCIS to IDC progression. Our preliminary observations lead to central hypothesis that tumor size-induced hypoxia establishes a “hypoxic secretome”, which initiates the migratory phenotype; the hypoxic secretome then cooperate with intracellular signaling networks to independently maintain cell migration. We propose three independent but inter-related aims to link hypoxic secretome with the initiation, maintenance and spatial distribution of migratory phenotypes. Aim 1 will engineer size-controlled DCIS organoids (150-600 µm) with controlled hypoxic microenvironments to identify and examine how hypoxic secretome initiates migratory phenotype. We will combine experimental organoid models with time-lapse imaging and computational approaches to study organoid migration. Aim 2 will demonstrate that migratory cells can re-establish the secretome and maintain migratory phenotype independent of hypoxia. We will reconstruct an intracellular signaling network activated by the hypoxic secretome using microarray data. We will verify these gene expression signatures in sorted migratory and non-migratory cells, and validate them using secretome inhibition studies. Aim 3 will investigate, for the first time, the spatial distribution and origin of the migratory phenotype. We will use CRISPR-based gene knock-in (FP-labeling), automated image analyses, and a deep-learning algorithm to track and visualize the emergence of migratory phenotypes from the hypoxic core outward to the periphery or from the migratory front. The successful development of this 3D organoid model and completion of the proposed work will provide answers to two fundamental questions in the progression of invasive breast cancer: 1) What causes some DCIS cells to become migratory and develop into invasive tumors? 2) How and where does the migratory phenotype (IDC) emerge? The mechanistic understanding gained from these studies will improve diagnosis, lead to the development of treatment strategies to arrest invasion at the pre-malignant stage, and thus prevent patient overtreatment. It is straightforward to generalize our system to other tumor types, development of tumor/stromal co-culture, and drug screening.
Title: A New Molecular Mechanism to Bioengineering a Liver
Description: Hepatocyte transplantation has many potential applications. Extensive animal experiments have shown that hepatocytes transplanted in the liver or at ectopic sites survive, function, and actively participate in the regenerative process. However, our understanding of hepatocyte engraftment and their remarkable proliferative and regenerative potential is limited, even if primary hepatocyte transplantation is at the doorstep of applications in the treatment of inherited and acquired human diseases. We previously made a serendipitous observation that normal hepatocytes transplanted in the peritoneal cavity of an animal with lethal liver disease migrate into the lymphatic system and engineer ectopic liver-like organoids that rescue an animal model from a fatal metabolic disorder. How hepatocytes enter the lymphatics and what molecular mechanism is responsible for the generation of ectopic mass is not known. We hypothesized that hepatocytes must borrow some of the molecular mechanism lymphocytes use to migrate into the lymphatics. Our interest will be to study ectopic cell transplantation and our central objective of our application is to translate a highly interesting observation, the generation of ectopic liver, to a potential clinical application for patients with liver diseases.
Description: The facility at 2124 Penn Ave. in Pittsburgh’s Strip District includes office and laboratory space where biotechnology companies can collaborate. In addition to the physical space, the LifeX platform brings together experts with proven track records of building new pharmaceuticals, medical devices, molecular diagnostics and population-health solutions to support entrepreneurs with deep industry knowledge. LifeX’s initial cohort of companies will focus on unmet health needs related to cancer, Alzheimer’s disease, multidrug-resistant bacterial infections, obesity and diabetes and rare genetic diseases.
Description: Researchers plan to spend the first few years of the grant improving or adding to existing standards, particularly related to cushion load-bearing performance, cushion durability, caster durability, and wheel rolling resistance, and the remaining few years applying those standards to different products. Ultimately, the researchers envision their work being used in several ways, including helping funding sources make reimbursement decisions, and clinicians, providers, and users make product decisions.
Description: The researchers will compare spinal manipulation and supported self-management to usual medical care, which includes prescription medications for the care of acute lower back pain in adult patients at increased risk of becoming chronic.
Description: Physical mechanical processes are central to the morphogenesis of embryos and their organs. The goal of this proposal is to apply a multi-scale analysis of the mechanics of convergent extension, identifying biomechanical mechanisms that establish passive tissue properties such as stiffness as well as active processes that generate forces of extension, regulate cell behaviors and tissue deformation, and how passive mechanics and active force generating processes are coordinated within the frog embryo. Studies outlined in this proposal will answer: (1) How are cell-scale structures and tissue mechanics are integrated during elongation? Early development is marked by dramatic changes in the mechanical properties of embryos. To understand how and why these properties change we test simple models of tissue mechanics based on synthetic closed-cell foams using bioengineering and biophysical methods to disrupt features from large scale architecture to the subcellular actomyosin-dependent cortex. (2) What single-cell mechanical processes contribute to convergent extension? We extend our analysis of cell behaviors to an unbiased approach that combines wide-field confocal microscopy with descriptive biomechanical analyses from the level of the cell, to the local neighborhood, to the strain fields of the entire embryo. Combining analyses of neural plate and paraxial somitic mesoderm we describe the dependence of these movements on planar polarity signaling. Using systems approaches we seek to test the dependencies of specific cell behaviors on both upstream signaling systems and their targeted downstream effectors. (3) How are tissue polarity cues transduced into polarized cell behaviors? We hypothesize that polarized cell behaviors and the oriented forces they generate are the result of cues produced by anisotropic strain. To test the roles of polarized intracellular factors and mechanical strain in organizing cell behaviors we use magnetogenetics and micro-scale tissue stretchers. Results from this project will complement ongoing efforts to identify the molecular regulators of morphogenesis by providing a conceptual framework developing new hypotheses of morphogenesis and bioengineering tools to test them. The significance of our work provides researchers a more complete understanding of the contribution of cell- and tissue-mechanics to development, to understand the role of tissue mechanics in oncogenesis, and to provide fundamental physical principles for future functional tissue engineers.
Description: This technology is a new ex-vivo application for sonothrombolysis (SNT), which combines the use of ultrasound (US) probes and microbubbles timely infused through the arterial port of liver allografts being preserved by a machine perfusion (MP) system. The US probe pulses induces microbubble oscillation and bursting in a process called cavitation. This ex-vivo technology is intended to remove red blood cells (RBCs) plugs and cellular debris from the hepatic arterial peri-biliary plexus (PBP) prior to liver allograft implantation from organs obtained from donors after cardiac death (“DCD”) These patients experience extended periods of hypoperfusion under anoxic conditions prior to organ recovery. The use of DCD livers poses a significant risk for the subsequent development of ischemic cholangiopathy (IC) by the recipient in the post-operative period. IC is an irreversible complication stemming from prolonged ischemia to the PBP leading into recurrent biliary sepsis and subsequent liver allograft failure. This lethal condition requires mandatory retransplantation while yielding prolonged hospital stays and excessive post-operative costs. Previous attempts to prevent IC after DCD liver transplantation using different technologies have failed. IC is caused by progressive clotting of the small blood vessels supplying the PBP, which prevents blood and oxygen from reaching the biliary tree effectively once the liver is transplanted. The ex-vivo SNT technology was designed to remove the clots ex-vivo while enhancing the oxygenation of the bile duct system before the liver is transplanted. It can be used with all current MP systems currently being evaluated for liver preservation.
Title: 3D Bioprinted Human Trachea for Pediatric Patients
Description: The overall goal of this project is the development and preclinical testing of a tissue engineered trachea for use in pediatric patients. The natural growth of pediatric patients requires that an engineered tissue or organ vital to life must “grow” with the patient. The present project will design, develop, and test in preclinical models, a bioengineered trachea consisting of naturally occurring extracellular matrix (ECM), as a scaffold, that is custom manufactured by 3D printing (Feinberg Laboratory, Carnegie Mellon University). The combined expertise of the Badylak Laboratory, which will acquire and prepare the matrix materials, with the Feinberg Laboratory that has expertise in 3D printing, will produce engineered tracheas that will be tested in a rapidly growing porcine model at the McGowan Institute for Regenerative Medicine. The project is milestone driven and consists of two years of development followed by three years of testing.
PI: William Federspiel, William Wagner, Peter Wearden
Title: Ambulatory Assist Lung for Children
Description: Acute and chronic lung diseases remain the most life threatening causes of death and hospitalization in the pediatric population. Cystic fibrosis (CF), pulmonary hypertension and pulmonary fibrosis have been observed to be the most frequent causes of lung failure in pediatric patients. Mechanical ventilation (MV) and extracorporeal membrane oxygenation (ECMO) have been used to bridge sick kids to transplant. These procedures can lead to poor post-transplant outcomes by their very restrictive nature on mobility. This project will develop a compact respiratory assist device for pediatric patients, the Pittsburgh Pediatric Ambulatory Lung (P-PAL) to replace ECMO as a bridge to transplant or recovery in kids with acute and chronic lung failure. The P-PAL is a wearable and fully integrated blood pump and gas exchange module that will be designed for implantation of inflow cannula and outflow cannula/grafts in the right atrium and pulmonary artery, respectively. The P-PAL will be designed for longer-term respiratory support (1-3 months before cartridge change-out) at 70-90% of normal metabolic oxygenation requirements, while pumping blood from 1 to 2.5 Liters/min. The specific aims of project are 1) To modify the design and operational parameters of the P-PAL to meet requirements for blood pumping, gas exchange, priming volume, and form factor, 2) To build P-PAL prototypes along the design development pathway for bench characterization studies of pumping performance, gas exchange, and hemolysis, 3) To improve the hemocompatibility of the P-PAL by exploiting novel polymeric zwitterionic coatings that we have already begun to develop for our adult wearable assist lung, and 4) To perform acute and chronic studies in healthy lambs to demonstrate the in-vivo performance and hemocompatibility of the PAAL device and to study its interaction with the cardiopulmonary system.
Title: Enhanced Neural Prosthetics Using Shared-Mode Control
Description: This project builds on the world’s most advanced program in brain-controlled robotic arms and hands for paralyzed individuals. A group of Pittsburgh scientists and engineers will enhance the performance of neural prosthetics, allowing a paralyzed person to manipulate an object. A prosthetic limb will operate under an individual’s brain control, with a boost from an artificial intelligence component designed to predict what the individual intends to do. This shared-mode control will enable people who have quadriplegia to dexterously handle objects with a robotic arm and hand, and thus increase their independence in daily life.
Title: Endotypes of thrombocytopenia in the critically ill
Description: Thrombocytopenia is extremely frequent in critically ill patients. However, the role of acute platelet responses in critically ill patients is not well studied, and the multifactorial etiology of thrombocytopenia in the ICU makes it difficult to understand, or understand whether or not to treat it. In several situations such as traumatic injury or sepsis, very low platelet counts have been to bleeding, thrombosis and end-organ injury. Platelets have been extensively studied as a key component of hemostasis, but a rapidly emerging concept is that platelets are also key effector cells in systemic inflammatory processes as both instigators of local and systemic inflammatory reactions and also participants in the inflammation that contributes to tissue injury. The link between platelets and inflammation is complex and bidirectional, as inflammatory ligands have been shown to regulate platelet function and activated platelets induce inflammatory responses in other cell types. The overarching theme of this proposal is to study platelet dynamics in critically ill patients, construct clinical endotypes of thrombocytopenia in this population, and to relate these endotypes to underlying mesoscale mechanisms through computational modeling. We will use a large electronic health record-based database and a tri-state trauma database as source data to construct these endotypes. We define endotype as clinical patterns defined along four dimensions: (1) baseline information (demographic, chronic disease burden, severity of illness and admitting diagnosis), (2) features of the platelet count time series (rate of decrease, nadir, etc.), (3) concurrent interventions, and (4) outcome. The computational approach will attempt to root clinical endotypes in mechanistic interpretations (or collections of alternative interpretations), contributing to focus basic science investiagtions, and to close key knowledge gaps preventing the design and use of targeted anti-platelet-inflammatory therapies in the critically ill. Computational models will be developed at different levels of complexity, with a specific attention to tie underlying mechanisms to functional assays routinely performed in thrombocytopenic patients, such as prothrombin time, activated coagulation time, and thromboelastogram.
Description: PUMP is a solution aimed at reducing hospital-acquired pressure ulcers, affecting an estimated 3 million patients annually. The monitoring and alert solutions, using wearable devices and hospital bed sensors, will provide real-time documentation of patient repositioning and a process to improve compliance with these preventative measures.
Title: IPA#14 – Development and Evaluation of Xenografts for Soft Tissue Reconstruction; IPA#15 – Development and/or Evaluation of Synthetic Materials, Synthetic/Biologic Material Composites, surgical hemostats/sealants/adhesives and/or Methods for Improving the Host Tissue Response to Such Materials; IPA#16 – Development and/or Refinement of In Vitro Methods which would Characterize and/or Predict the Host Response to a Test Article; and IPA#17 – Development / Refinement of Preclinical Models and Ex-Vivo Test Methods
Title: Outside-In Regenerative Therapy for Abdominal Aortic Aneurysm
Description: Few diseases represent the optimal potential target for regenerative cellular therapy more than the abdominal aortic aneurysm (AAA). A disease that affects a large number of elderly in the United States with a natural history that results in structural failure of the aortic wall and death, AAA continues to represent a critical need for biologic therapy. Regenerative therapy consisting of the delivery of stem cells to the damaged aorta presents a conceptually strong opportunity for the reconstitution of the aortic mural matrix and therefore aortic strength – any test of such a therapy must be done on an established aneurysm to most accurately represent what occurs in the clinic. In this proposal, we have combined the strengths of two laboratories with complementary scientific capability, and with a common interest in the development of effective biologic therapies for AAA disease. The early product of this collaborative pairing is published “proof of concept” evidence that mesenchymal stem cell (MSC) delivery to the wall of a murine AAA can slow progression. The purpose of this R21 Exploratory/Developmental Research Proposal is to develop a clinically- translatable MSC delivery system that would result in aortic matrix repair and regeneration. Our hypothesis is that local stem cell delivery to a murine AAA via an adventitially-applied hydrogel and magnetic assistance will result in intramural cell engraftment, matrix repair, and mechanical stabilization of the aortic wall. To address our hypothesis, we will execute the following specific aims: Specific Aim 1 is to develop and validate a technique to deliver MSCs into the aortic wall periadventitially using a hydrogel vehicle and magnetic guidance. The technique will be optimized by testing a cadre of iron nanoparticle types, fibrin hydrogel formulations, and stem cell concentrations both in vitro and in vitro. Specific Aim 2 is to demonstrate that local MSC delivery halts and reverses the functional and structural degeneration of an AAA in an established rodent model. MSC hydrogels developed in Specific Aim 1 will be applied to the adventitia of an elastase-induced model AAA after allowing for varying degrees of matrix degeneration. Metrics for success of the various therapies versus cell-free hydrogel controls on aortic tissue will involve: i) functional assessment (including aortic diameter and biomechanical parameters) and ii) detailed microstructural and cellular composition assessment. The expected outcome of this work is the development and proof-of-concept of a new technology for stem cell delivery to AAA.
PI: Michael L. Boninger, MD and Thomas A. Rando, MD
Title: Alliance for Regenerative Rehabilitation Research & Training (AR3T)
Description: The advancement of regenerative medicine principles and technologies holds great potential to drive progress in the prevention and treatment of individuals with a host of pathologies resulting from injury, disease or aging. The long-term goal of regenerative medicine is to promote the repair, replacement, or regeneration of tissues. Likewise, rehabilitation seeks to harness the body’s innate regenerative potential in order to maximize function. Both fields hold great potential to drive progress in the treatment of a host of acute and chronic pathologies. We propose that these two fields are inextricably intertwined; an intersection of disciplines known as Regenerative Rehabilitation. To fully realize the tremendous potential of Regenerative Rehabilitation, we must promote the interaction of basic scientists with rehabilitation specialists. We must also train rehabilitation clinicians who can help oversee the quality, safety, and validity of these innovative Regenerative Rehabilitation technologies. The overarching goal of the Alliance for Regenerative Rehabilitation Research & Training (AR3T) is to establish a national network that will expand scientific knowledge, expertise and methodologies across the domains of regenerative medicine and rehabilitation.
Description: Sepsis, a clinical systemic inflammatory response syndrome occurring in patients following infection or injury, remains the leading cause of death in intensive care units worldwide, including the United States. Emerging evidence indicates that immunometabolism may play an important role in the pathogenesis of sepsis through its ability to regulate the expression and release of cytokines. In particular, we recently provided the first direct evidence that PKM2-mediated aerobic glycolysis promotes the release of HMGB1, a late mediator of lethal systemic inflammation with a wider therapeutic time window for clinical intervention. These exciting findings raise several important questions regarding the previously unknown role of PKM2 in the pathogenesis of sepsis, as well as the novel mechanisms underlying the regulation of PKM2 expression and HMGB1 release. We hypothesize that PKM2-mediated immunometabolism is an emerging hallmark of sepsis that contributes to cytokine (e.g., HMGB1) release and the subsequent systemic inflammatory response. We propose the following specific aims:
Title: Extracellular Matrix as a Therapy for Inflammatory Bowel Disease
Description: Clinical Problem: Inflammatory Bowel Disease (IBD) affects nearly 1.5 million Americans with approximately 70,000 new cases diagnosed each year. There are a number of forms of IBD most notably ulcerative colitis (UC) and Crohn’s disease, but the medical device therapy described in this work could also address proctitis, proctosigmoiditis, and rectal mucositis. UC is the initial research target that may or may not include the other forms of inflammatory disease in the colon. UC significantly increases the risk of developing colorectal cancer and negatively impacts quality of life. The etiology of UC is unknown but altered intestinal barrier function and production of cytotoxic effector molecules within the mucosal lining of the colon are known to play central roles in the development of UC. The proposed work would investigate and evaluate the ability of a hydrogel form of extracellular matrix (ECM) to restore structure and function of the mucosal lining of the colon in a preclinical UC model. These findings would represent an important contribution to required regulatory filings prior to human studies.