Investigators - Basic science - shankland

Stuart J. Shankland, MD, MBA

Project 1: Glomerular epithelial cell regeneration and progenitors

Podocytes are terminally differentiated epithelial cells who’s function include limiting the passage of albumin from the intravascular space, to the extravascular urinary space. However, injury in several primary proteinuric glomerular diseases such as FSGS and secondary glomerular diseases such as diabetes, oftentimes results in podocyte depletion due to events such apoptosis, detachment and altered autophagy. In contrast to other kidney cells, podocytes do not have the cell cycle machinery to adequately proliferate. Thus, renewal of podocyte number in adults much rely on the regeneration by progenitor cells. Our laboratory is studying two potential podocyte progenitor cells: cells of renin lineage that reside in the juxta-glomerular compartment, and glomerular parietal epithelial cells, more immediate neighbors for podocytes. Using a number of specific reporter mice, we can show that following the depletion of podocyte number in experimental models of glomerular disease, a subset of cells of renin lineage move to the intraglomerular compartment, where they begin to express markers of podocytes, as well as acquire the characteristic ultrastructure of these highly specialized cells. A subset of cells of renin lineage in the glomerulus also begin to express markers of parietal epithelial cells Taken together, our research shows that both cells of renin lineage and parietal epithelial cells serve a role as podocyte progenitors. Studies are ongoing to determine the underlying pathways and mechanisms, and ways in which regeneration can be augmented, as the ultimate goal is to improve the outcomes of patients with glomerular disease.

Project 2: Cell cycle proteins in podocyte biology

Our earlier work focused on the more “classic” cell cycle regulatory proteins, where we showed that following injury to podocytes, levels of specific cyclin-dependent kinase inhibitors (p21, p27, p57) increased, thereby inhibiting the activity of cyclin-cyclin dependent kinase complexes. This limits and prevents podocyte proliferation, leading to an overall depletion in cell number. More recently, we reported that podocytes constitutively express cyclin I and cdk5, considered more “atypical” cell cycle proteins. Our data shows for the first time that cyclin I activates cdk5, and that active cyclin I-cdk5 is essential for podocyte survival following stress in disease. This novel biological role is governed by c ertain MAPK pathways which in turn regulate critical pro-survival Bcl-2 family proteins. We are exploring other pathways whereby cdk5 is activated, including p35, and how the interplay of cyclin I and p35 might determne the subcellular localization and thus function of their partner cdk5.

Project 3: Role of pericytes in tubulo-interstitial scarring

Together with Dr. Jeremy Duffield, we are continuing a body of work showing that kidney pericytes lead a dual life. On one hand they are vital for normal peri-tubular vascular function, where they surround these vessels and maintain their survival. Yet, under certain disease-induced stress situations, pericytes become more fibroblast-like, and become pro-fibrotic cells. This leads to progressive interstitial fibrosis and a decrease in kidney function.

Project 4: Diabetic kidney disease

Together with Dr. Charles Alpers, our laboratory is exploring the consequences of reduced podocyte number, and how these cells might be replaced through regeneration by progenitor cells. In experimental type 2 diabetic nephropathy, we recently reported that parietal epithelial cells might be a candidate podocyte progenitor, and studies are ongoing using specific reporter mice to address this hypothesis.

Current Funding

NIH – R01- DK083391 Shankland (PI) (4/1/2011-3/31/16)
Podocyte depletion and regeneration during evolution and reversal of diabetic nephropathy
This grant explores mechanisms underlying reversibility (replacement of kidney cell populations that are typically lost in human and experimental diabetic kidney disease) with emphasis on the podocyte, a unique cell type in the kidney. This grant also focuses on testing new therapies, based on correcting metabolic injuries to mitochondria (an organelle present within all cells), that are specifically directed to podocyte mitochondria, as a way to promote reversal of diabetic kidney disease.

American Heart Association 11GRNT7810021 Shankland (PI) (7/1/2011-6/30/13)
Cell Cycle Regulation of Podocyte Proteins
The overall goal of this grant proposal is to prove that both slit diaphragm and actin-binding proteins are Regulated by the cdk5-cyclin I-p35 complex, and that any alterations in this complex such as occurs in disease, leads to proteinuria and glomerulosclerosis.

NIH-RO1 Shankland (PI) (7/1/10 - 6/30/15)
Podocyte Progenitor Cells
The focus of this grant application is to delineate how terminally differentiated podocyte cell number is Restored following injury in glomerular disease. Based on our recent data and preliminary data, we have compelling evidence that renin-expressing juxtaglomerular cells and parietal epithelial cells both serve as local progenitor cells. We will show that they both switch their phenotype in disease, and ultimately become podocytes.

NIH-RO1- Renewal Shankland (PI) (7/1/11 - 6/30/16)
Cell Cycle and Podocyte Apoptosis
Studies are designed to determine the role of Cyclin I and CDK5, and when active, this complex enhances Survival of terminally differential cells such as podocytes. The signaling pathways will be delineated. Cell culture and experimental models will be used to delineate this further.

Past Funding

NIH ARRA Grant (DK056799-07S1) (09/21/09-08/31/11)
Cell Cycle and Podocyte Apoptosis
Studies are designed to determine the role of Cyclin I and CDK5, and when active, this complex enhances survival of terminally differential cells such as podocytes. The signaling pathways will be delineated. Cell culture and experimental models will be used to delineate this further.

NIH R01 Grant (DK056799-05A1) Shankland (PI) (09/30/07 – 6/30/11)
Cell Cycle and Podocyte Apoptosis
The overall goal of this grant is to delineate new paradigms in the regulation of podocyte survival and death, so that ultimately new strategies can be developed to prevent podocyte loss, enhance kidney survival, and reduce kidney disease.

NIH Basic Training Grant (T32 DL07467-21) Shankland(PI) (09/01/2009–8/31/13)
Nephrology Training 03
This is a training grant that supports research training for three M.D. post-graduate fellows who undergo periods of both renal and basic science research training.

NIH NIDDK ARRA R21 (DK081835-01A2) Shankland (PI) (08/15/09 – 06/30/11)
New Thoughts on Parietal Cells
The precise biological role of parietal epithelial cells (PECs) are not well understood in health or disease. The main goal of this grant is to show that PECs are required to limit filtered proteins ‘escaping’ into the peri-glomerular space. We will test the hypotheses that the intra-cellular tight junctions in PECs, together with the underlying Bowman’s Basement membrane, forms a second barrier to proteinuria. We will also test the hypothesis that PECs actively take up filtered albuminuria which injures them by inducing apoptosis.

1 F32 DK072788-01 Shankland (PI) (9/19/05-12/30/07)
HHS Logar Fellow

DHHS 5901 Letter of Credit (LOC) Shankland (PI) (4/1/2008-3/31/09)
7th Intl Podocyte Conference
This grant will be used to support the travel of junior investigators to attend scientific conference.

AHA – Established Investigator Award 0340129N Shankland (PI) (1/01/03– 12/31/08)
Role of Cell Cycle in Podocyte Injury
The proposed studies are designed to test novel aspects of cell cycle proteins in disease, so that potential therapeutic interventions can be designed to reduce the incidence of renal failure.

NIH R01 grant DK 51096-05A2 Shankland (PI) (09/01/04-06/30/07)
Cell Cycle Control in Glomerular Disease
The major goal of this grant is to determine the expression and activity of specific cell cycle proteins in glomerular cell proliferation and differentiation in vitro and in vivo. Overlap: None.

Applied Molecular Genetics (AMGEN) Shankland (PI) (09/27/04-09/26/06)
Role of Erythropeitin (EPO) in Proteinuria
The overall goal of this grant is to test whether a decrease in endogenous EPO in renal disease augments loss of podocytes differentiation, preventing progressive proteinuria and scarring.

American Diabetes Association (7-04-RA-107-80-0572) Shankland (PI) (07/01/04-06/30/06)
Role of Podocytes in Diabetic Nephropathy
The overall goal of this grant is to delineate the mechanisms underlying the abnormalities in podocytes, utilizing cultured cells and animal models of diabetic nephropathy.

NIH R01 grant DK 60525-02 Shankland (PI) (07/01/02-06/30/06)
Mechanisms of Podocyte Injury
The major goals of this grant are to study the effect of stress-tension-induced injury on podocytes, and also to determine the role of novel cell cycle regulatory proteins in podocyte disease.

NIH R01 grant DK 56799-04 Shankland (PI) (05/01/00-02/28/05)
Cell Cycle Proteins and Glomerular Apoptosis
The major goal of this project is to show novel roles for specific cell cycle proteins in glomerular cell apoptosis beyond that of proliferation.

1-2001-395 Shankland (PI) (01/01/01-12/31/03)
Juvenile Diabetes Research Foundation (JDRF)
The Role of Cyclin Kinase Inhibitors in Diabetic Glomerular Hypertrophy
The major goal of this project will be to determine the nuclear mechanisms underlying the development of diabetic hypertrophy, with a focus on specific CDK inhibitors. Overlap: None.

NIH R01 grant DK 47659-10 Shankland (PI) (09/01/98-08/31/03)
Immunological Renal Diseases: Subproject #2, “The Role of Cell Cycle Proteins in Glomerular Epithelial Cell Growth”
The major goal of this project is to test the hypothesis that specific CDK-inhibitors p57-WT1 determine the proliferative capacity of the glomerular visceral epithelial cell.

NIH R01 grant DK 51096 Shankland (PI) (06/01/96-05/31/04)
Cell Cycle Control in Glomerular Disease
The major goal of this grant is to determine the expression and activity of specific cell cycle proteins in glomerular cell proliferation and differentiation in vitro and in vivo. Overlap: None.

NIH/NIDDK R13 (DK069139-01) Shankland (PI) (06/01/04-06/30/05)
“5th International Podocyte Conference”
This grant was used to support the travel of junior investigators to attend this scientific meeting.

NIH (R01) Grant; (DK 52121) Shankland (PI) (01/01/97-12/31/01)
Role of the Glomerular Endothelial Cell in Hemolytic Uremic Syndrome (HUS)
The goal was to determine the mechanisms of endothelial cell proliferation and apoptosis.

C. V. Therapeutics, Inc. Grant Shankland (PI) (01/01/00-12/31/00)
Role of Cell Cycle Proteins in Hemodialysis Vascular Stenosis
The study was designed to determine the role of cell cycle proteins in vascular access stenosis.

Current Research Members

Diana Eng, PhD: Research Scientist
Diana Eng received her BS in Bioengineering at the University of Washington in 2008 and her PhD in Molecular and Cellular Biology at Oregon State University in 2013. She joined the Shankland Lab as a Research Scientist upon graduation and is currently working on identifying and characterizing the molecular mechanisms of kidney regeneration.

Natalya Kavarina, MD, PhD: Research Scientist
Dr. Natalya Kaverina is a trained scientist-physician, who obtained her PhD degree from the N.N. Blokhin Russian Oncology Research Center, Moscow, Russia. She then completed her training at the laboratory of Lupus Research, Department of Rheumatology, University of Chicago prior to joining the Dr’s Shankland laboratory, Department of Nephrology, University of Washington. Natalya has extensive experience with multicolor immunofluorescence and immunohistochemistry, by using those techniques, she characterized different subsets of lymphocytes in human lupus nephritis. Her current area of research focuses on the biology of cells of renin lineage in normal and diseased states. To test the hypothesis that cells of renin lineage serve as progenitors to replace lost podocytes in disease, Natalya is using the inducible multi-reporter (confetti) mice, activated by the renin gene.

Julia Lichtnekert, MD, PhD: Research Fellow
Julia graduated from the Ludwig Maximilian University Munich, Germany in 2008 after finishing medical school. Since 2006 she has been studying the expression and function of innate immune receptors in inflammatory kidney diseases. During the medical thesis Julia investigated inflammatory effects of necrotic cell death in primary mesangial cells. She contributed to studies investigating the role of viral RNA and DNA inducing type I interferons in mesangial cells. Later she concentrated on the role of inflammasome signaling in intrinsic glomerular inflammation and studied the contribution of resident renal cells including renal dendritic cells in IL-1β induction in the kidney. During her clinical residency Julia studied the therapeutic effects of activated protein C in a mouse model of systemic lupus erythematosus and lupus nephritis. She continued her previous studies on immune mechanisms in inflammatory kidney diseases in the laboratory of Dr. Duffield in Seattle. To further extend her focus to repair and regeneration processes in kidney injury Julia joined the Shankland lab in January 2014.

Bairbre McNicholas, MD, PhD: Research Fellow
With a background in biochemistry, medicine, and nephrology, Bairbre’s clinical interests include diabetic kidney disease, hyperkalemia, and urinary biomarkers. During her PhD studies, she studied intra-renal inflammation in obesity and type 2 diabetes mellitus. In Dr. Shankland’s lab, she is currently studying the role of cells of renal lineage in mouse models of type 1 and type 2 diabetes mellitus.

Jeff Pippin: Senior Laboratory Scientific Director/Research Scientist and Lab Manager
Jeff obtained Bachelor's of Arts in Biology from Southern Illinois University in 1989. He worked at Washington University in St. Louis from 1989-1992 in Rheumatology studying the role of arachidonic acid metabolites in glomerulonephritis. After moving to Seattle, he worked for Fred Hutchinson Cancer Research Center from 1992 to 1993 in Pediatric Oncology. He then joined the University of Washington, Division of Nephrology in 1993 working with Dr. William Couser, studying the role of complement in glomerular disease. He began working with Dr. Stuart Shankland in 1996, initially studying the cell cycle in experimental models of kidney disease, followed by a focus on podocyte biology and most recently glomerular repair and regeneration. Jeff has extensive of expertise in experimental rodent models of kidney disease, kidney cell culture, cell biology and pathology, with over 90 authored and co-authored publications.

Ania Stefanska, PhD: Research Fellow
Ania obtained her MSc degree in biotechnology from the Jagiellonian University in Krakow, Poland where she was working on muscle regeneration. Following a student exchange in Finland she developed an interest in vascular biology. She was awarded a British Heart Foundation-funded PhD in cardiovascular biology in Edinburgh, Scotland, UK. Her PhD project concerned primarily pericytes in human kidney development and their ability to act as renin progenitors in vitro. In Dr. Shankland’s lab Ania will be investigating renin lineage cells in the context of kidney injury/regeneration.

Publications

PubMed provides a list of Dr. Shanklandi's research publications

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