Journal of Orthopedics & Rheumatology
Searching for the Molecular Pathways Regulating Bone Mineral Density in the Proteome and RNA Interference Era
Fawzy A. Saad*
- Department of Orthopaedic Surgery, Albert Einstein College of Medicine, USA
*Address for Correspondence: Fawzy A. Saad, Department of Orthopaedic Surgery, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA, Fax: +1 718 430 3259; E-mail: firstname.lastname@example.org
Citation: Saad FA. Searching for the Molecular Pathways Regulating Bone Mineral Density in the Proteome and RNA Interference Era. J Orthopedics Rheumatol. 2013;1(1): 7.
Copyright © 2013 Saad FA. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Journal of Orthopedics & Rheumatology | ISSN: 2334-2846 | Volume: 1, Issue: 1
Submission: 23 October 2013 | Accepted: 04 December 2013 | Published: 09 December 2013
Reviewed & Approved by: Dr. Ming Pei, Department of Orthopedics, West Virginia University, USA
AbstractOsteoporosis is a polygenic disorder associated with low bone mineral density and deterioration of bone microarchitecture with increased chance of bone fractures. Although bone matrix mineralization and osteoporosis are closely related, the mineralization of bone matrix is almost a forgotten aspect in osteoporosis research. The complex processes of bone matrix mineralization and bone remodeling are tightly regulated by several transcription factors and signal transduction pathways. However, signal transduction pathways occurring at a protein level that depends not only on mRNA transcriptional regulation but also on a multitude of translational and posttranslational controls. Furthermore, proteomics allow a discerning view of complex molecular pathways, provides an efficient method to determine protein candidates, and elucidates signal transduction pathways that regulate bone mineral density and accelerates the discovery of osteoporosis causative genes. RNA interference is a powerful tool for rapid analysis of gene functions. Therefore, strategies to combine proteomics with RNA interference and transgenic RNAi would greatly improve the efficiency of gene discovery and divulge the molecular pathways involved in osteoporosis pathophysiology. In this review, current methods employed to identify genes involved in osteoporosis, which include linkage analysis, candidate gene association studies, genome wide association studies, transcriptome microarray, and proteomics are evaluated, and a new strategy is proposed.
KeywordsMesenchymal stem cells; Osteoblast matrix mineralization; Bone mineral density; Bone metabolic disorder; Bone remodeling; Bone tissue engineering; Bone gene therapy; Osteoporosis; Genome; Transcriptome; Proteome; Linkage analysis; Proteomics-2RNAi
AbbreviationsDlx5: Distal-less Homeobox 5; ATF4: Activating Transcription Factor 4; SATB2: Special AT-Rich Sequence-Binding Protein 2; Twist1: a basic helix-lop-helix transcription factor; MITF: Microphthalmia-Associated Transcription Factor
IntroductionCompletion of the human genome project more than a decade ago holds great promise for scientific research to excavate the genetic foundation of complex biological processes such as bone matrix mineralization. However, bone matrix mineralization is nearly a forgotten dimension in osteoporosis research . Osteoporosis is a polygenic disorder determined by multiple genes and environmental risk factors, each with modest effects on bone mass and susceptibility to fracture. It is a bone metabolic disorder associated with low Bone Mineral Density (BMD) and deterioration of bone microarchitecture [2,3] with increased chances of bone fracture. BMD changes with age, having a rapid increase during the childhood to reach a peak level by the mid or late twenties in life and declining thereafter in women and elderly men, which is due to unbalanced bone remodeling. Bone matrix mineralization is an important determinant of the stiffness and hardness of the bone material [4,5]. Moreover, it has become evident in recent years that bone mineral and matrix tissue properties play a pivotal role in the overall biomechanical competence of bone [6,7]. The process of bone matrix mineralization is tightly regulated both temporally and spatially . Some factors, such as mineral-binding-extracellular matrix proteins and proteoglycans, mineralization-inhibiting proteins and matrix-vesicles [9,10], are known but still very little is known about the molecular control of bone matrix mineralization. An enhanced understanding of the regulatory mechanisms underlying bone matrix mineralization may improve our understanding of the molecular basis of osteoporosis pathophysiology.
Suitable for identifying gene responsible for monogenic disorders
Lack the sensitivity to identify genes underlying polygenic disorders.
Candidate genes association studies
Several candidate genes in a signaling pathway may be studies simultaneously.
Inconsistent, spurious, and insignificant replication of the association study results.
Genome wide association studies
Suitable to investigate genetic architecture of polygenic disorders arising from nucleotide polymorphisms. Offers the possibility to identify novel susceptibility genes and pathways.
It does not identify individual causal genes, nor does it provide functional information required for discovering a therapy, and the occurrence of false negatives is highly significant.
Offers insights into the global patterns of gene expression, and provides a panoramic analysis of gene expression alterations.
Signal transduction occurs at protein level, and the correlation between mRNA and protein abundance in the cell is extremely poor.
An efficient method to determine protein candidates, and elucidates signal transduction pathways that regulate bone mineral density and accelerates the discovery of osteoporosis causative genes.
Still under development and show certain limitations, which include the capabilities to identify challenging protein groups such as low-abundance, hydrophobic and basic proteins.
Methods for Identifying Osteoporosis GenesIn the proteome era, diverse methods have been currently employed to identify genes involved in complex genetic disorders, which include linkage analysis, candidate gene association studies, genome wide association studies, transcriptome microarray, and proteomics.
Main Biological functions
High density lipoprotein
RNA binding protein
Phosphoglycerate kinase 1
Tooth germ development
Protein disulfide isomerase A3
Pentose phosphate pathway
Prolyl 4-hydroxylase a1
Prolyl 4-hydroxylase a2
Pyruvate kinase muscle
MITF transcription activities
Actin binding protein
New StrategyProteomics is a promising approach to increase understanding about the molecular pathways which underlie the complex process of bone matrix mineralization, and offers the prospective to overcome the limitations of the genome and transcriptome based approaches. Moreover, proteomics provides an efficient way to elucidate the signal transduction pathways regulating bone mineral density. The WNT pathway is a key regulator of skeletogenesis and osteoblast differentiation. Meanwhile, Receptor Activator of NF-Kappab (RANK) ligand pathway regulates osteoclasts and bone homeostasis. Both WNT and RANK ligand signalling pathways pathways are linked to osteoporosis [98-100].
ConclusionsGenetic factors play important roles in the development of osteoporosis, but the genes and mutations conferring osteoporotic risk remain largely unknown. Linkage analysis, first developed to map gene alterations causing monogenic bone disorders like osteogenesis imperfecta, appear to lack the sensitivity to define genes underlying polygenic bone disorders. Until now, few causative genes have been discovered and most of the genetic variables leading to osteoporosis remain to be identified, which is mainly because linkage analysis lacks the sensitivity to map genes responsible for polygenic disorders. The conclusive failure of linkage analysis to identify causative genetic factors in polygenic disorders has led scientists to focus on candidate gene association studies.
AcknowledgementsThe author would like to thank Melvin J. Glimcher for his continuous support, and Natalie B. Saad for reading the manuscript.
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