Identification of novel bone-specific molecular targets of binge alcohol and ibandronate by transcriptome analysis.

paper Cited Public

Authors: Himes, Ryan; Wezeman, Frederick H; Callaci, John J
Year: 2008
Journal: Alcoholism, clinical and experimental research
PMID: 18537941
DOI: 10.1111/j.1530-0277.2008.00736.x
PMCID: PMC2728683

BACKGROUND: Our laboratory established that binge alcohol-related bone damage is prevented by aminobisphosphonates, suggesting bone resorption increases following binge exposure. We examined the effects of binge alcohol and antiresorptive therapy on the relationship between bone damage and modulation of the vertebral transcriptome, in an attempt to determine how alcohol-induced bone damage and its prevention modulate bone-related biological pathways. METHODS: Male Sprague-Dawley rats were assigned to 1 of 6 treatment groups (n = 12/group). (C1) saline ip 3 d/wk for 1 week, (A1) binge alcohol, 3 g/kg, ip 3 d/wk for 1 week, (C4) saline ip, 3 d/wk for 4 weeks, (A4) binge alcohol, ip, 3 g/kg 3 d/wk for 4 weeks, (I4) ibandronate, saline ip 3 d/wk for 4 weeks, plus a single ip injection of ibandronate at 120 microg/animal, and (AI4) binge alcohol plus ibandronate as above. After 1 or 4 weeks, adjacent lumbar vertebrae were assayed for bone damage or transcriptional changes. RESULTS: Bone loss was not observed after 1 week of binge alcohol treatment. After 4 weeks, binge alcohol decreased vertebral BMD by 23% (p < 0.05) and compressive strength by 18% compared to saline controls (p < 0.05). Concurrent ibandronate prevented bone loss, increasing these parameters by 145 and 134% respectively compared to binge alcohol. (p < 0.05). Analysis of the vertebral transcriptome identified gene clusters specific for acute and chronic binge alcohol-related bone damage. Acute binge alcohol modulated the expression of integrin signaling-specific genes, while chronic binge alcohol modulated canonical Wnt signaling gene expression. Ibandronate normalized the expression of approximately 20% of the genes affected by chronic binge alcohol, allowing the identification of a unique subset of alcohol-sensitive, ibandronate-responsive genes. CONCLUSIONS: Identification of bone-specific gene expression clusters associated with acute and chronic binge alcohol treatment allowed for the identification of cellular pathways affected by binge treatment with known involvement in bone remodeling (Integrin, Canonical Wnt signaling) not previously identified as alcohol-sensitive. This data provides a basis for a plausible mechanistic explanation for the known detrimental effects of alcohol on bone formation and resorption.

#	Section	Preview
20	RESULTS — Effects of Acute and Chronic Binge Alcohol Exposure on the Vertebral Transcriptome	study. We identified 1300 annotated differentially expressed genes in bone after an acute binge…
21	RESULTS — Effects of Acute and Chronic Binge Alcohol Exposure on the Vertebral Transcriptome	observed which sort into the pathway over the number of genes expected based on a reference list.…
22	RESULTS — Effects of Acute and Chronic Binge Alcohol Exposure on the Vertebral Transcriptome	Chronic binge alcohol exposure pathway-related effects are shown in Fig. 4. Significant effects were…
23	RESULTS — Gene Clusters Associated With Acute and Chronic Binge Alcohol-Related Bone Damage	Quality Threshold (QT) clustering was used to identify a gene cluster from the acute binge alcohol…
24	RESULTS — Gene Clusters Associated With Acute and Chronic Binge Alcohol-Related Bone Damage	Similarly, the list of genes differentially expressed only after a chronic binge alcohol cycle was…
25	RESULTS — Identification of a Binge Alcohol-Ibandronate Signature Profile	We next searched our expression dataset to determine if a gene cluster could be identified that…
26	RESULTS — Identification of a Binge Alcohol-Ibandronate Signature Profile	superimposed with a bar graph showing the effects of chronic binge alcohol on vertebral cancellous…
27	RESULTS — Validation of Gene Array Data by Real-Time RT-PCR	Real-Time quantitative RT-PCR (qRT-PCR) analysis was used to confirm the direction and magnitude of…
28	DISCUSSION	This study utilized a young adult rat model of binge alcohol exposure, which mimics the pattern and…
29	DISCUSSION	The current study shows a clear association between the duration of binge alcohol exposure and the…
30	DISCUSSION	determine relative contributions of body weight versus treatment on measured parameters found no…
31	DISCUSSION	Gene array analysis was a tool utilized to gain an understanding of the biology behind the effects…
32	DISCUSSION	Although no significant effects of acute binge alcohol on vertebral BMD or strength were observed,…
33	DISCUSSION	of a significant number of genes involved in integrin signaling, including a significant increase in…
34	DISCUSSION	Chronic binge alcohol-related differential expression also had significant effects on cellular…
35	DISCUSSION	β-catenin (ctnnb1), and Dickkopf-1 (Dkk1). β-catenin, referred to as the molecular node of the…
36	DISCUSSION	Another alcohol-related effect on bone formation potentially explained by a deactivation of…
37	DISCUSSION	Quality Threshold (QT) clustering, which allowed the visualization of all existing patterns of gene…
38	DISCUSSION	and has not been previously reported. This observation is supported by a previous unrelated study…
39	DISCUSSION	In summary, the data presented in this investigation suggests that alcohol-related bone damage may…

Name	Type
1700-microarray analyzer local	drug
A1 local	cohort
A1 treatment group local	cohort
A4 local	cohort
A4 treatment group local	cohort
ABI chemiluminescence detection kit local	drug
ABI chemiluminescent RT-IVT labeling kit local	drug
ABI microarray analyzer software package local	drug
ABI rat genome survey microarray local	drug
Acute binge alcohol local	drug
Acute Binge Alcohol Exposure Treatment Group local	cohort
Acute binge alcohol treatment local	cohort
Acute binge alcohol treatment local	phenotype
acute binge exposure local	drug
Adipocyte local	phenotype
Agilent 2100 Bioanalyzer	drug
AI4 local	cohort
AI4 treatment group local	cohort
AI treatment group local	cohort
alcohol	phenotype
Alcohol-modulated bone gene cluster local	gene
Alcohol plus ibandronate (AI) treatment group local	cohort
Alcohol-related bone damage local	phenotype
alcohol-related bone loss local	phenotype
Alcohol-related bone loss local	phenotype
Alcohol-related osteopenia local	phenotype
alcohol withdrawal	phenotype
animals	cohort
Applied Biosystems 1700 Gene Expression Array System local	drug
B2M local	gene
Binge Alcohol local	drug
Binge alcohol and ibandronate-treated local	cohort
Binge alcohol-treated local	cohort
Binge Alcohol Treated Animals local	cohort
binge alcohol treatment local	cohort
binge alcohol treatment local	drug
binge alcohol treatment local	phenotype
Bisphosphonate local	drug
blood alcohol level	phenotype
blood alcohol levels	phenotype
BMD	phenotype
body weight	phenotype
body weight increase local	phenotype
Bone architecture local	phenotype
Bone cement local	drug
bone damage local	phenotype
Bone Damage local	phenotype
Bone formation local	phenotype
bone loss	phenotype
Bone mass local	phenotype
Bone Metabolism local	phenotype
bone mineral density	phenotype
Bone mineral density (BMD) local	phenotype
Bone Remodeling local	phenotype
bone resorption	phenotype
bone strength local	phenotype
Bone strength local	phenotype
C1 local	cohort
C4 treatment group local	cohort
Cancellous BMD local	phenotype
Cancellous bone mineral density local	phenotype
Canonical Wnt signaling pathway local	drug
cDNA	drug
Chemokine/cytokine signaling local	phenotype
Chondrocyte local	phenotype
chronic binge alcohol local	drug
Chronic binge alcohol local	drug
Chronic binge alcohol group local	cohort
chronic binge alcohol-treated rats local	cohort
Chronic binge alcohol-treated rats local	cohort
Chronic binge alcohol treatment local	cohort
Chronic binge alcohol treatment local	drug
Chronic Binge Alcohol Treatment Group local	cohort
CO2	drug
Compressive strength local	phenotype
Compressive Strength local	phenotype
Condition	phenotype
control animals	cohort
control group	cohort
control rats	cohort
Cortical bone mineral density local	phenotype
Crohn's disease	phenotype
CTNNB1	gene
Digoxigenin-UTP-labeled cRNA local	drug
DKK1	gene
Estrogen-dependent bone loss local	phenotype
ethanol consumption	phenotype
Fatty bone marrow local	phenotype
female rats	cohort
Fifth lumbar vertebra local	phenotype
food intake	phenotype
Fourth lumbar vertebra local	phenotype
fragmentation buffer	drug
genes	gene
High bone mass phenotype (HBM) local	phenotype
hybridization buffer local	drug
ibandronate local	drug
Ibandronate local	drug
Ibandronate-treated local	cohort
integrin	drug
Integrin signaling local	phenotype
integrin signaling genes local	gene
Itgav local	gene
Itgb3 local	gene
ITGB3 local	gene
L3 vertebrae local	phenotype
L4 vertebral body local	phenotype
L5 vertebral body local	phenotype
LRP5	gene
LRP6 local	gene
lumbar vertebrae local	phenotype
male rats	cohort
Mesenchymal stem cells (MSC) local	phenotype
NAD+	drug
Nanodrop	drug
Osteoblast local	phenotype
Osteopenia local	phenotype
osteoporosis	phenotype
Osteoporosis pseudo-glioma syndrome (OPPG) local	phenotype
PDGF signaling local	phenotype
post-treatment weight local	phenotype
Ras signaling local	phenotype
rats	cohort
RiboPure RNA isolation kit local	drug
RNA	drug
RNase H	drug
saline	drug
Saline control local	cohort
Sprague-Dawley rats	cohort
TNFRSF11B	gene
total RNA	drug
treatment group	cohort
TUBB	gene
vertebral BMD local	phenotype
Vertebral BMD local	phenotype
vertebral body area local	phenotype
Vertebral bone local	phenotype
Vertebral cancellous BMD local	phenotype
Vertebral compression fractures local	phenotype
vertebral compression strength local	phenotype
vertebral compressive strength local	phenotype
Vertebral cortical BMD local	phenotype
vertebral strength local	phenotype
vertebral trabecular BMD local	phenotype
Young adult rat model local	cohort

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In this knowledge base

Title	Year	PMID
Genome-wide association study of alcohol dependence implicates a region on chromosome 11.	2010	20201924

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