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Current Molecular Medicine

Volume 5, Number 5, 2005

 

Contents

 

Diseases of the Kidney: Molecular Mechanisms and Current Therapy

Executive Editor: Stephen I-Hong Hsu

 

Editorial Pp.453-454

Stephen I-Hong Hsu

[Abstract]

 

New Developments in the Field of Cystic Kidney Diseases Pp.455-465

Ralph Witzgall

[Abstract]

 

Cellular and Molecular Pathways that Lead to Progression and Regression of Renal Fibrogenesis Pp.467-474

Hirokazu Okada and Raghu Kalluri

[Abstract]

 

Molecular Basis of IgA Nephropathy Pp.475-487

Andrew S.H. Lai and Kar Neng Lai

[Abstract]

 

Bone and the Kidney: A Systems Biology Approach to the Molecular Mechanisms of Renal Osteodystrophy Pp.489-496

Adrian Mondry, Zhengyuan Wang and Pawan K. Dhar

[Abstract]

 

Genetic and Genomic Approaches to Glomerulosclerosis Pp.497-507

A. Padiyar and J.R. Sedor

[Abstract]

 

Molecular Genetic Approaches for Studying the Etiology of Diabetic Nephropathy Pp.509-525

D.P.K. Ng and A.S. Krolewski

[Abstract]

 

Conditional Gene Targeting in the Kidney Pp.527-536

Alexander Gawlik and Susan E. Quaggin

[Abstract]

 

Novel Non-rodent Models of Kidney Disease Pp.537-546

Dirk M. Hentschel and Joseph V. Bonventre

[Abstract]

 

Abstracts

 

[Back to top] Editorial

Stephen I-Hong Hsu

 

The pandemic of kidney disease in the 21st century affects increasing numbers of individuals and is associated with significant morbidity and mortality as well as a heavy economic burden. It has been long predicted that this pandemic will wreck its greatest havoc in highly populated developing nations such as China and India, along with the remainder of Asia/Southeast Asia, where the incidence of type 2 diabetes mellitus is on the rise. Clearly, as a world wide clinical and research community, nephrology is in a state of emergency. Something must be done—and soon.

 

Although we continue to be challenged by our incomplete understanding of the molecular pathophysiology of acute and chronic renal diseases, including genetic diseases involving the kidney, it is safe to say that there has never been a more exciting or important time to be a “molecular nephrologist.” Historically, it may be argued that nephrology, as a scientific discipline, has been slower than its sister disciplines such as cardiology and endocrinology, to adopt both the conceptual and technical approaches and platforms offered by the revolution in molecular biology begun in the early 80’s. Apologetics notwithstanding, the stunning success of renal anatomists and physiologists during the “pre-molecular” era, and which continues to this day, may have contributed to the prolonged pregnancy and delayed birth of modern molecular nephrology. If this is indeed the case, then the contents of this volume of current reviews—representing only a small but representative sampling of the activity of a dedicated and exceptionally gifted community of modern molecular nephrologists engaged in a broad spectrum of scientific activities—are a testament to the coming-of-age of molecular nephrology in the post-genomic era.

 

It is perhaps worth considering for a moment the value of the category of the review article and its function in science. At minimum, review articles succinctly catalogue the latest developments in a specialized area of investigation, often directly or indirectly helping the reader to appreciate areas of controversy or pointing out a critically important gap in knowledge that has been inadvertently ignored. At its best, a review article may stand as a genuine contribution to the body of published primary data, when the author is able to offer a new direction or propose a novel theory or model based on a synthesis of published data. In this manner and only judged in retrospect, a review article may prove to be highly influential. Although the authors of the reviews in this volume were simply instructed to name any topic and any title of their own choosing, and despite the obvious differences in the styles adopted, it is enormously gratifying to find that each has produced a review article that may be acknowledged as having achieved a level of the very highest order.

 

What better example of the above than Witzgall’s review of cystic kidney disease? After managing to concisely cover important new developments and insights arising from the study of a rather broad category of diseases, he shows us how the application of molecular methods has unexpectedly uncovered a common theme (the role of the primary cilium at the apical membrane of renal tubular epithelial cells) and has brought us to refocus attention on a fundamental and unsolved problem in basic cell biology research—the “problem of geometry.” He speculates that primary cilia and their associated proteins play a role in determining proper tubular geometry. Similarly, Okada and Kalluri emphasize the importance of delineating the molecular pathways of fibrogenesis that underlie progressive chronic kidney disease (CKD)—a common clinical outcome irrespective of the primary injury or disease process. They speculate that genomic approaches hold the promise for revealing new therapeutic targets that will lead to novel strategies to prevent and/or reverse renal fibrosis. Lai and Lai provide a highly comprehensive review of a large body of work related to the molecular characterization of the immunopathogenesis of a fascinating and elusive disease, IgA nephropathy, the most common primary glomerulonephritis worldwide for which no widely accepted therapy or non-invasive diagnostic test is currently available. They highlight a series of important recent and novel findings upon which rational therapeutic strategies can begin to be designed and implemented. The first section is rounded out by a review of the syndrome of renal osteodystrophy that logically argues in favor of the appropriateness of a novel systems biology approach towards a more dynamic and quantitative understanding of the physiology and pathophysiology of renal bone disease.

 

While the first section can be loosely labeled as “disease-centric,” the second section highlights the availability and proper application of powerful molecular genetic approaches to elucidating molecular pathogenesis in both human and animal models. This is done in the context of a particular disease or disease process. Padiyar and Sedor take us through recent studies of familial focal segmental glomerulosclerosis to demonstrate how genetic and genomic approaches such as linkage analysis that have led to the identification of mutations in protein components of the slit diaphragm in rare causes of glomerulosclerosis (known collectively as podocytopathies), have led to novel insights regarding the general function of the slit diaphragm in health and disease. The authors should be complimented for their clearly stated assessment of the dismal state of population-based genetic association studies of candidate genes for CKD. The review by Ng and Krolewski reiterates the paradigm of molecular genetic approaches for identifying genetic variants that make strong (linkage analysis) and modest or weak (association studies) contributions to the phenotype of diabetic nephropathy. Importantly, they build on this paradigm by demonstrating the relative advantages of population- versus family-based association studies, and by emphasizing the potential synergy between human and experimental animal approaches (predominantly in mouse). They provide excellent examples of well-designed linkage and association studies that are ideally suited to serve as templates for the investigator who would venture into the potentially treacherous waters of molecular genetic studies. These two reviews provide a natural segue to the last two reviews that focus on powerful non-human experimental models for the study of kidney disease and normal renal physiology. Gawlik and Quaggin provide a highly readable, authoritative and up-to-date review of conditional gene targeting in the kidney, illustrating the newly available tools for achieving both spatial and temporal control of gene expression in the mouse. Recently published techniques for tissue-specific gene knockdown using RNA interference, which can theoretically be applied in a genome-wide, high throughput and cost effective manner, as well as a discussion of evolving forward genetic techniques such as random mutagenesis in mice, are also covered. It is clear that a tissue-specific conditional gene knockout in mouse will become a widely applied gold standard for defining the function of a specific gene. Hentschel and Bonventre provide the first published journal review of novel non-rodent models for the study of kidney disease, emphasizing the advantage of non-rodent genetic models for which the entire sequence of the genome has been determined. They nicely contrast the relative advantages of studying small organisms in which large scale screening approaches are feasible and cost-effective, as compared to larger mammals in which physiological and pathophysiological responses more closely mimic that of humans (especially useful in transplantation immunology studies). They also provide the images that grace the cover of this special issue of Current Molecular Medicine, which are taken from a recently published novel zebrafish model of acute renal failure.

 

I wish to thank the authors for the generous gift of their contribution of time and expertise, without which this issue would not have been possible. Special thanks to Anil Mukherjee (Editor-in-Chief) for granting me carte blanche on this project. Happy reading!

 

[Back to top] New Developments in the Field of Cystic Kidney Diseases

Ralph Witzgall

 

For quite some time the field of polycystic kidney disease has led a life at the fringe of kidney research, but with the cloning of the PKD1 and many other genes this situation has dramatically changed. Polycystic kidney disease often is a syndromic disease affecting a variety of organs in addition to the kidney. Most of the proteins involved in polycystic kidney disease have been localized to the primary cilium, an extension at the apical membrane of renal tubular epithelial cells, which may serve chemo- and mechanosensory functions. It is speculated that primary cilia and their associated proteins play a role in determining the proper tubular geometry.

 

[Back to top] Cellular and Molecular Pathways that Lead to Progression and Regression of Renal Fibrogenesis

Hirokazu Okada and Raghu Kalluri

 

Renal fibrosis is a common consequence and often a central feature of all the progressive renal diseases that lead to end-stage renal failure. In comparison to wound healing, during kidney fibrosis the length of the post-inflammatory phase often exceeds and continues unchecked resulting in scar formation. Infiltrating immune cells and a heterogeneous colony of interstitial cells derived from a variety of cellular origins such as resident mesenchymal cells, tubular epithelial cells, circulating fibrocytes, and bone marrow derived stem cells, communicate with each other and with inflamed and surviving parenchymal cells via a network of cytokines and adhesion molecules to populate the renal tubulointerstitial space during early fibrogenesis. Such fibroblasts subsequently secrete abundant extracellular matrix to achieve architectural remodeling in parallel with functional deterioration. Renal fibrosis is a dominant determinant of the clinical outcome of patients and yet for the most part, current therapies are ineffective or only marginally effective. This review highlights recent advances in our understanding of the cellular and molecular events leading to the progression of renal fibrosis.

 

[Back to top] Molecular Basis of IgA Nephropathy

Andrew S.H. Lai and Kar Neng Lai

 

IgA nephropathy (IgAN) is the most common glomerulonephritis worldwide and remains an important cause of end-stage renal failure. However, the basic molecular mechanism(s) underlying abnormal IgA synthesis, selective mesangial deposition with ensuing mesangial cell proliferation and extracellular matrix expansion remains poorly understood. Notably, the severity of tubulointerstitial lesions better predicts the renal progression than the degree of glomerular lesions. The task of elucidating the molecular basis of IgAN is made especially challenging by the fact that both environmental and genetic components likely contribute to the development and progression of IgAN. This review will summarize the earlier works on the structure of the IgA molecule, mechanisms of mesangial IgA deposition and pathophysiologic effects of IgA on mesangial cells following mesangial deposition. Recently, a series of important advances in the area of communication between the glomerular mesangium and renal tubular cells have emerged. These novel findings regarding the molecular pathogenesis of IgAN will be helpful in designing future directions for therapy.

 

[Back to top] Bone and the Kidney: A Systems Biology Approach to the Molecular Mechanisms of Renal Osteodystrophy

Adrian Mondry, Zhengyuan Wang and Pawan K. Dhar

 

Despite its apparent static condition, the skeleton undergoes a permanent process of remodeling mediated by osteoblasts and osteoclasts. The activity of these cells is regulated by a plethora of factors, ranging from mechanical stress to the effects of hormones to the immune system. One well-studied regulatory system involves the maintenance of calcium homeostasis through a network whose main regulatory components include ionized calcium, phosphate, parathyroid hormone and active vitamin D. This system establishes the link between bone and kidney, as one of the kidney’s endocrine functions is the activation of vitamin D, while electrolyte homeostasis is one of its excretory functions. Impaired renal function leads to disturbances in this regulatory system, resulting in the complex syndrome of renal osteodystrophy that affects the majority of patients with chronic renal failure. This review summarizes the current understanding of bone physiology on a molecular level, examines some of the pathological pathways related to renal disease, and concludes with an outlook on how the emerging field of systems biology may contribute to a more dynamic and quantitative understanding of the physiology and pathophysiology of renal bone disease.

 

[Back to top] Genetic and Genomic Approaches to Glomerulosclerosis

A. Padiyar and J.R. Sedor

 

Chronic kidney disease (CKD) is common, progressive and expensive to manage. Although modifiable risk factors can be treated and outcomes improved, CKD remains a chronic disease with excessive morbidity and mortality. The completion of the human genome sequence and the advent of methodologies to define gene function provide new opportunities to manage and treat patients with CKD and other chronic diseases. Despite the lack of clear correspondence between genotype and phenotype and an obvious Mendelian inheritance pattern, CKD susceptibility has a genetic basis. In this review, we focus on recent studies of familial focal segmental glomerulosclerosis and the discoveries that have resulted from both genetic and genomic approaches used to understand its pathogenesis. Key slit diaphragm proteins were discovered using linkage analyses of these rare causes of glomerulosclerosis and subsequent work has characterized slit diaphragm function in health and disease. Podocyte dysfunction is now recognized as a key contributor to the functional and histologic derangements that characterize glomerular dysfunction in many common causes of CKD. In aggregate, these studies provide a paradigm for approaches to better define mechanisms of CKD and to identify novel therapeutic targets.

 

[Back to top] Molecular Genetic Approaches for Studying the Etiology of Diabetic Nephropathy

D.P.K. Ng and A.S. Krolewski

 

A critical challenge faced by clinical nephrologists today is the escalating number of patients developing end stage renal disease, a major proportion of which is attributed to diabetic nephropathy (DN). The need for new measures to prevent and treat this disease cannot be overemphasized. To this end, modern genetic approaches provide powerful tools to investigate the etiology of DN. Human studies have already established the importance of genetic susceptibility for DN. Several major susceptibility loci have been identified using linkage studies. In addition, linkage studies in rodents have pinpointed promising chromosomal segments that influence renal traits. Besides augmenting our understanding of disease pathogenesis, these animal studies may facilitate the cloning of disease susceptibility genes in man through the identification of homologous regions that contribute to renal disease. In human diabetes, various genes have been evaluated for their risk contribution to DN. This widespread strategy has been propelled by our knowledge of the glucose-activated pathways underlying DN. Evidence has emerged that a true association does indeed exist for some candidate genes. Furthermore, the in vivo manipulation of gene expression has shown that these genes can modify features of DN in transgenic and knockout rodent models, thus corroborating the findings from human association studies. Still, the exact molecular mechanisms involving these genes remain to be fully elucidated. This formidable task may be accomplished by continuing to harness the synergy between human and experimental genetic approaches. In this respect, our review provides a first synthesis of the current literature to facilitate this challenging effort.

 

[Back to top] Conditional Gene Targeting in the Kidney

Alexander Gawlik and Susan E. Quaggin

 

Complete mapping of the genome in a number of organisms provides a challenge for experimental nephrologists to identify potential functions of a vast number of new genes in the kidney. Since knockout technologies have evolved in the early eighties the mouse has become a valuable model organism. Researchers can now artificially eliminate the expression of specific genes in a mammalian organism and examine the phenotype. New developments have emerged that allow investigators to knock out a gene specifically in the kidney. Several kidney-specific promoters provide valuable tools and bacterial artificial chromosome (BAC) based techniques like recombineering will enhance both number and accuracy of new mouse lines with spatially controlled gene expression. In addition to spatial control, tetracycline- or tamoxifen-inducible systems, provide the possibility of influencing the temporal expression pattern of a gene enabling researchers to dissect its functions in adult organisms. Knocking out a gene will continue to be the gold standard for defining the role of a specific gene whereas tissue-specific gene knockdown using RNA interference represents an alternative approach for generating lower-priced and fast loss of function models. In addition to reverse genetic approaches, forward genetic techniques like random mutagenesis in mice continue to evolve and will enhance our understanding of disease mechanisms in the kidney.

 

[Back to top] Novel Non-rodent Models of Kidney Disease

Dirk M. Hentschel and Joseph V. Bonventre

 

Kidney disease in the 21st century affects increasing numbers of individuals. We continue to be challenged by our lack of understanding of the pathophysiology of acute and chronic renal disease including genetic diseases involving the kidney. Rodent knockout animals or inbred strains have greatly contributed to our understanding of many monogenetic and complex diseases. Non-rodent animal models of disease have become more attractive since genomic data has become available for a variety of organisms that offer distinct advantages over mice and rats for ease in conducting high-throughput chemical or mutagenesis screens. It is thus timely to examine the physiology and pathophysiology of the kidney or kidney equivalents in these organisms to evaluate their relevance as models for human disease. In addition to organisms whose small size and accessibility facilitate large scale screening approaches, larger animals at the other end of the spectrum offer unique physiological advantages in both size equivalency to humans as well as, in some cases, physiological and pathophysiological responses that closely mimic those of humans. Here we review a selected number of non-rodent experimental models of kidney diseases, focusing on recent advances in the use of the worm Caenorhabditis elegans, the fruitfly Drosophila melanogaster, the zebrafish Danio rerio, the little skate Leucoraja erinacea, the MGH miniature swine, merino cross sheep, and the cow Bos taurus to study kidney disease.