Bone and cartilage physiology
Research in our group is aimed at understanding the molecular mechanisms underlying the physiological regulation of bone and cartilage homeostasis.
Andrei Chagin – Forskare
Jussi Heinonen – Anknuten
Lei Li – Doktorand
Phillip Newton – Anknuten
Baoyi Zhou – Doktorand
Björn Barenius, MD, PhD, Head of Section for Arthroplasty and Arthroscopy, Department of Orthopaedic Surgery, Södersjukhuset.
- Eva Lindgren (research engineer)
- Giuseppe Di Lernia (master student, Karolinska Institutet)
- Rinat Gizatulin (visiting researcher)
- Sima Rahbar Zare, (master student, University of Skövde, Sweden, 2012-2013)
- Oskar Ljungkvist (summer student, Karolinska Summer School, 2012)
- Frida Kozma (summer student, Karolinska Summer School, 2012)
- Amanda Demir (summer student, Karolinska Summer School, 2013)
- Klas Walldén (summer student, Karolinska Summer School, 2013)
- Savas Ay (medical student from Dicle University, Turkey, supported by Erasmus program, summer 2013)
Project 1: G-protein pathways involved in the growth plate fusion
Humans do not grow during the entire length of their life-time. We stop growing due to fusion of the so-called growth plate or epiphyseal plate - a thin layer of chondrocytes located between the epiphysis and the metaphysis of long bones. Fusion of the growth plate normally occurs during puberty and results in cessation of longitudinal growth. However, mechanisms of the growth plate fusion are completely unknown.
It was recently shown that ablation of the parathyroid hormone (PTH) receptor (PTHR1) in chondrocytes induces fusion of growth plate in mice. The main mediator of PTHR1 action is the alpha-subunit of the stimulatory G-protein (Gsa). However, we found that Gsa does not mediate fusion of the growth plate entirely and there are additional g-proteins involved in this process. It was reported that Gq/11 and Ga12/13 g-protein families can be activated by PTH.
To test the role of g-proteins in the growth plate fusion we genetically inactivate g-proteins in chondrocytes and monitor bone growth and behavior of the growth plate.
These experiments will reveal factors involved in the growth plate fusion. The obtained knowledge could lead to the development of new drugs, targets and strategies regulating fusion of the growth plate. This, in turn, can help children with growth abnormalities, for example extremely tall stature.
Please see KI News: New findings explain why we stop growing at one point
Project 2: Role of autophagy in longitudinal bone growth, chondrocyte survival and hypertrophy
The main endocrine regulator of longitudinal growth is the growth hormone/insulin-like growth factor (GH/IGF1) system. In many cellular systems IGF1 is signaling via PI3K/Akt/mTOR pathway. Mammalian Target of Rapamycin (mTOR) kinase is the main regulator of autophagy. Autophagy is promoted primarily by inhibition of mTOR, followed by phosphorylation of the Atg proteins responsible for autophagosome formation. Autophagy is a cellular process of catabolic degradation of damaged organelles and long-lived protein complexes. In addition to providing building blocks and energy, autophagy is involved in heart hypertrophy, neurodegeneration, immunological responses and bone turnover. A recent genome-wide association studies demonstrated that the regulation-of-autophagy pathway has the strongest association with bone size.
We hypothesize that autophagy plays an important role in chondrocyte physiology, contributing to cell survival and chondrocyte hypertrophy, as well as participating in endocrine and nutritional regulation of bone growth.
To explore the contribution of autophagy to longitudinal bone growth we utilize both ex vivo and in vivo models. For ex vivo experiments we use our recent model of postnatal mouse bone cultures. We treat bones with pharmacological modulators of autophagy and monitor their growth in vitro.
To modulate activity of autophagy in mice in a time- and tissue-specific manner we are inactivating either Atg5 or Atg7 genes utilizing Cre-lox approach. Inactivation of either Atg5 or Atg7 abrogates autophagy. We employ the following transgenic mice: Atg5 floxed, Atg7 floxed, cartilage specific Collagen-2-driven Cre (Col2-Cre), and tamoxifen-inducible Col2-Cre. Upon breeding Cre and floxed strains, progenies have no autophagy specifically in their chondrocytes. We thereafter characterize the growth plate phenotype of the obtained mice and their responsiveness to hormones and nutrition.
Understanding of mechanisms regulating longitudinal bone growth could lead to development of new drugs to modulate linear growth for children with growth disorders.
Project 3: Mechanisms of cell size increase
Children’s long bones grow from narrow cartilage organs located at the ends of the bones called growth plates. Cells in the growth plate, called chondrocytes, increase in size by up to fifteen times their volume which determines 60-80 % of overall bone growth. Very little is known about the mechanisms within the chondrocytes, which controls this cellular enlargement, called hypertrophy, either at the basic research or at the clinical level. This situation is emphasized by the fact that the usual treatment for children suffering from extreme tall or short stature is invasive surgery or hormone therapy, respectively. These procedures are suboptimal, expensive, and moreover they do not target the cause of the problem directly. If we are to treat these patients more rationally and efficiently, further research of the downstream signalling pathways coordinating growth plate development is needed. Our research goal is to discover how chondrocyte hypertrophy is coordinated at the cellular level. This project has the possibility to uncover the molecular mechanisms and therefore reveal pharmaceutical approaches to treat children with extremely tall or short stature, which would vastly improve the current treatments.
Project 4: Osteoarthritis
Osteoarthritis is an age-dependent degradation of articular (joint) cartilage. This is a very common problem affecting more than 50 % of Swedish people above 65 years of age. Etiology of this disease is poorly known, with many potential underlying mechanisms including genetics, trauma, aging etc.
In our laboratory we are exploring mechanisms of cartilage protection and underlying causes of osteoarthritis development.
A radical switch in clonality reveals a stem cell niche in the epiphyseal growth plate.
Newton PT, Li L, Zhou B, Schweingruber C, Hovorakova M, Xie M, et al
Nature 2019 03;567(7747):234-238
Targeted deletion of Atg5 in chondrocytes promotes age-related osteoarthritis.
Bouderlique T, Vuppalapati KK, Newton PT, Li L, Barenius B, Chagin AS
Ann. Rheum. Dis. 2016 Mar;75(3):627-31
Pharmacological inhibition of lysosomes activates the MTORC1 signaling pathway in chondrocytes in an autophagy-independent manner.
Newton PT, Vuppalapati KK, Bouderlique T, Chagin AS
Autophagy 2015 ;11(9):1594-607
Targeted Deletion of Autophagy Genes Atg5 or Atg7 in the Chondrocytes Promotes Caspase-Dependent Cell Death and Leads to Mild Growth Retardation.
Vuppalapati KK, Bouderlique T, Newton PT, Kaminskyy VO, Wehtje H, Ohlsson C, et al
J. Bone Miner. Res. 2015 Dec;30(12):2249-61
G-protein stimulatory subunit alpha and Gq/11α G-proteins are both required to maintain quiescent stem-like chondrocytes.
Chagin AS, Vuppalapati KK, Kobayashi T, Guo J, Hirai T, Chen M, et al
Nat Commun 2014 Apr;5():3673
See also news article on KI News: New findings explain why we stop growing at one point
Mice depleted of the coxsackievirus and adenovirus receptor display normal spermatogenesis and an intact blood-testis barrier.
Sultana T, Hou M, Stukenborg JB, Töhönen V, Inzunza J, Chagin AS, et al
Reproduction 2014 Jun;147(6):875-83
Dexamethasone differentially regulates Bcl-2 family proteins in human proliferative chondrocytes: role of pro-apoptotic Bid.
Zaman F, Chrysis D, Huntjens K, Chagin A, Takigawa M, Fadeel B, et al
Toxicol. Lett. 2014 Jan;224(2):196-200
Resveratrol treatment delays growth plate fusion and improves bone growth in female rabbits.
Karimian E, Tamm C, Chagin AS, Samuelsson K, Kjartansdóttir KR, Ohlsson C, et al
PLoS ONE 2013 ;8(6):e67859
Genetic regulation of the growth plate.
Karimian E, Chagin AS, Sävendahl L
Front Endocrinol (Lausanne) 2011 ;2():113
Mechanisms of growth plate maturation and epiphyseal fusion.
Emons J, Chagin AS, Sävendahl L, Karperien M, Wit JM
Horm Res Paediatr 2011 ;75(6):383-91
Insulin-like growth factor-1 restores dexamethasone-induced heart growth arrest in rats: the role of the ubiquitin pathway.
Chrysis D, Chagin A, Sävendahl L
Hormones (Athens) ;10(1):46-56
Parathyroid hormone/parathyroid hormone-related protein receptor signaling is required for maintenance of the growth plate in postnatal life.
Hirai T, Chagin AS, Kobayashi T, Mackem S, Kronenberg HM
Proc. Natl. Acad. Sci. U.S.A. 2011 Jan;108(1):191-6
Expression of vascular endothelial growth factor in the growth plate is stimulated by estradiol and increases during pubertal development.
Emons J, Chagin AS, Malmlöf T, Lekman M, Tivesten A, Ohlsson C, et al
J. Endocrinol. 2010 Apr;205(1):61-8
Catch-up growth after dexamethasone withdrawal occurs in cultured postnatal rat metatarsal bones.
Chagin AS, Karimian E, Sundström K, Eriksson E, Sävendahl L
J. Endocrinol. 2010 Jan;204(1):21-9