Comprehensive proteomic analysis of mesenchymal stem cell exosomes reveals modulation of angiogenesis via nuclear factor-kappaB signaling

by Johnathon D. Anderson, Henrik J. Johansson, Calvin S. Graham, Mattias Vesterlund, Missy T. Pham, Charles S. Bramlett, Elizabeth N. Montgomery, Matt S. Mellema, Renee L. Bardini, Zelenia Contreras, Madeline Hoon, Gerhard Bauer, Kyle D. Fink, Brian Fury, Kyle J. Hendrix, Frederic Chedin, Samir El-Andaloussi, Billie Hwang, Michael S. Mulligan, Janne Lehtiö, Jan A. Nolta

February 2016

Multispectral optical tweezers for biochemical fingerprinting of CD9-positive exosome subpopulations

by Randy P. Carney, Sidhartha Hazari, Macalistair Colquhoun, Di Tran, Billanna Hwang, Michael S. Mulligan, James D Bryers, Eugenia Girda, Gary S. Leiserowitz, Zachary James Smith, and Kit S. Lam

March 2017

Billanna Hwang, MPH, DHSc

Research Scientist/PI-in-training, Department of Surgery (University of Washington)

Founder, WestCo Exosortium

Billanna (Billie) is currently a Research Scientist and baby PI under the direction/training of Dr. Michael S. Mulligan. Her research background is in bone marrow and solid organ transplant immunology with focus in tolerance induction/mechanisms, GVHD, and regulatory T cells. Majority of her research history has been at Fred Hutchinson Cancer Research Center and she continues to do collaborative research with those investigators. She continues her research using cell immunotherapy in pulmonary diseases and as a platform for tolerance induction in a solid organ transplant setting. In the last three years, exosomes have become a part of her research repertoire. This research includes 1) exo-immunomodulation, 2) exo-based therapy, 3) exo-profiling in patients, and 4) exo-regeneration.

Michael S. Mulligan, MD

Chief of Cardiothoracic Surgery (University of Washington), Director of the Lung Transplant Program, Director of Advanced Lung Disease Surgery Program

Dr. Mulligan is a UW professor of cardiothoracic surgery and the director of the Lung Transplant Program and the Advanced Lung Disease Surgery Program. He is clinically busy in lung transplantation and lung volume reduction surgery (LVRS). He also is the surgeon in the Pacific Northwest who performs pulmonary endarterectomies for chronic pulmonary embolic disease.

In addition, Dr. Mulligan directs the Minimally Invasive Thoracic Surgery Program. He performs a range of lung cancer resections regularly. He is considered a national expert in video-assisted thoracic surgery (VATS) lobectomy and leads regional and national courses on the subject. He also has considerable experience in the management of esophageal cancer.

He has won local and national teaching awards and runs an NIH-funded laboratory. His areas of scientific investigation include the mechanisms of acute lung injury following lung transplantation. regulation of tolerance in long-term lung graft acceptance, and lung ischemia reperfusion injury.

Thomas Anchordoquy, PhD

Professor, Department of Pharmaceutical Science (University of Colorado, Denver)

Dr. Thomas Anchordoquy has been working on lipid-based drug delivery systems for over 30 years, and he finds it interesting that nature has evolved a similar delivery system. In collaboration with Michael Graner, they have studied the ability of exosomes to deliver drugs in vitro and in vivo. In addition, they have done extensive proteomic analysis of exosomes harvested under various conditions. He would be interested in communicating with anyone who is exploiting exosomes or the exosomal pathway for drug delivery.

Johnathon Anderson, PhD

University of California, Davis

Dr. Johnathon Anderson directs the Exosome Team at the University of California Davis’ Institute for Regenerative Cures. This works involves investigating mesenchymal stem cell derived exosomes as potential therapeutics for the treatment of ischemic tissue diseases (ischemic retinopathy, PAD) and acute brain injuries (ischemic stroke, TBI). This approach uses a combination of genetic engineering and high throughput omics strategies, combined with functional in vivo studies.

James D. Bryers, PhD

Professor, Department of Bioengineering (University of Washington)

Adjunct Professor, Department of Chemical Engineering (University of Washington)

The Bryers research group is one of perhaps only two or three engineering-based research groups investigating the interaction of bacteria, immune cells, and biomedical implant materials. Thier research over the past 20 years has defined and quantified the biological and physical processes governing (1) the formation and persistence of microbial biofilms in biotechnological and medical systems, (2) control of macrophage phenotype at biomaterial interfaces, and (3) developed biomaterials that promote infection immunity. Current research activities are (1) developing anti-biofilm biomaterials, (2) creating biomaterials that promote immunotherapy and enhance vaccine efficiency, and (3) tissue regeneration by exosome engineering.

Randy Carney, PhD

Assistant Professor, Department of Biomedical Engineering (University of California, Davis)

Dr. Carney’s research focuses on developing new platforms for early stage cancer diagnosis by applying spectroscopic methods to characterize circulating exosomes and related extracellular vesicles (EVs). The Carney Lab (https://carneylab.faculty.ucdavis.edu) builds cutting-edge Raman spectroscopy and SERS tools to investigate the broad heterogeneity of EVs, with the goal of more sensitively and specifically identifying tumor-associated subpopulations. Projects include (1) single vesicle Raman analysis of EVs isolated from ovarian cancer patient biofluids obtained from the UCD Comprehensive Cancer Center, (2) chemical and materials synthesis of next-generation plasmonic SERS nanoprobes, and (3) microfluidic chip-based platforms for isolation of EVs from precious clinical samples.

Gagan Deep, PhD

Associate Professor of Cancer Biology (Wake Forest School of Medicine)

Michael Graner, PhD

Associate Professor, Department of Neurosurgery (University of Colorado)

Dr. Graner's research focuses on the immunology and biology of brain tumors. From a clinical perspective, he is interested in vaccine design and implementation, which includes the search for appropriate combinations of therapies to enhance immune responses or to downplay the role of tumor-induced immune suppression.

Elena Hsieh, MD

Assistant Professor of Immunology & Microbiology and Pediatrics (University of Colorado, Denver)

Dr. Elena Hsieh earned her MD degree from University of California San Francisco (UCSF) in 2008. She completed a residency in pediatrics at the University of California Los Angeles (UCLA) in 2011, and a fellowship in Allergy and Immunology at Stanford University in 2014. She continued her research and clinical work at Stanford University as an Instructor for an additional year. In 2015, Dr. Hsieh joined the faculty at the University of Colorado Denver School of Medicine, jointly affiliated with the Children’s Hospital of Colorado. Dr. Hsieh studies immune dysregulation in autoimmunity, primary immunodeficiency, and the overlap between the two.

Michael Olin, MD

Assistant Professor, Division of Pediatric Hematology and Oncology (University of Minnesota)

Dr. Olin’s scientific interest is defining the mechanism(s) of suppression inhibiting the ability to mount a tumoricidal response evident in the tumor draining lymph nodes and tumor environment and developed potential ways to modulate the immunosuppressive activity. Immunosuppressive tumor environment is a major hurdle for the immune system to overcome. Dr. Olin is focused on the development of inhibitors derived to overcome the suppressive tumor microenvironment.

Joseph Quinn, MD

Professor of Neurology (Oregon Health and Science University)

Joseph Quinn, MD, is a Professor of Neurology at Oregon Health and Science University, engaged in translational research in neurodegenerative diseases including Alzheimer's and Parkinson's Disease.  He has a longstanding interest in cerebrospinal fluid markers as surrogate outcome measures for clinical trials.  As Director of the OHSU Alzheimer's Center Biomarker Core, he manages a repository of CSF and plasma samples from patients and control subjects.

Julie Saugstad, PhD

Associate Professor (Oregon Health & Science University)

My research is focused on elucidating the molecular and cellular mechanisms of signal transduction in brain injury and neurodegeneration. We arefocused on the role of microRNAs as effectors of post-transcriptional regulators of gene expression in response to ischemia and neuroprotection, as well as for their clinical utility as biomarkers for Alzheimer's Disease in human cerebrospinal fluid. My lab uses several molecular and biochemical techniques, including proteomics, microarrays, real-time quantitative PCR, immunocytochemistry, immunoblots, RNA blots, in situ hybridization, RNAi, and enzymatic assays.

Aijun Wang, PhD

Assistant Professor & Co-Director of the Surgical Bioengineering Laboratory (University of California, Davis)

Dr. Aijun Wang is an assistant professor of surgery. He is Co-Director of the Surgical Bioengineering Laboratory at UC Davis School of Medicine. Dr. Wang was trained in biology at Tsinghua University, China, and had undergone postdoctoral training at UC Berkeley Department of Bioengineering and Berkeley Stem Cell Center with a postdoctoral fellowship from California Institute for Regenerative Medicine (CIRM). He is a faculty member of UC Davis since 2012. Dr. Wang's research goal is to develop novel technologies that combine stem cell engineering and biomaterial engineering to promote tissue regeneration. Recently, Dr. Wang’s group has started investigating stem cell derived exosomes for tissue regeneration applications. 

Christina Coughlan, PhD, FCP, SI, NRAEMT

Research Instructor (University of Colorado, Denver)

Prior to becoming a Senior Faculty Research Instructor in Dr. Potter’s laboratory, all of my experience-both research and teaching-has focused on gaining a better understanding of neurodegeneration. My specific skills, expertise and drive as a Pharmacologist and Biochemist are to translate basic science findings into therapies and biomarkers for disease, in particular Alzheimer’s disease. I have focused on examining; ER stress, protein misfolding, the role of amyloid in Alzheimer’s, the subcellular regulation and distribution of molecular players in the development of Schizophrenia, the role of chemokines as neuronal migration cues, sialylation as an important posttranslational regulator of neural cell adhesion molecules and the role of its modifications on memory formation. In the past two years I have spent considerable research effort optimizing exosome isolation, purification and characterization, with the intent of translating markers in the plasma into biomarkers for both the progression of Alzheimer’s and for understanding the molecular mechanisms behind the improvement observed in patients receiving leukine, a study initiated in our lab by Dr. Timothy Boyd and Dr. Huntington Potter which is now in human Phase II trials.