Nada Lukkahatai, PhD, RN
Co-authors : B. Majors, BS; S. Reddy, PhD; B. Walitt, MD; L. Saligan, PhD



To examine differential expression of genes from fatigued fibromyalgia women with different levels of pain and catastrophizing.
Does the gene expression profile different in fatigued fibromyalgia women with high and low pain severities?
Does the gene expression profile different in fatigued fibromyalgia women with high and low catastrophizing?

 Fibromyalgia (FM) is characterized by prolonged widespread muscle pain, profound fatigue, and sleep disturbance.
 An estimated 10 million Americans and 200-400 million adults worldwide suffer from FM.
 Prevalence: 3 times higher among women than men.
 5.5 million FM patients visit the ambulatory care per year.
 The etiology of the fibromyalgia is unknown.

1990 American College of
Rheumatology (ACR)
Criteria
1. History of chronic widespread pain
2. Pain in at least 11/18 tender point sites on digital palpation.

1 . Widespread Pain
Index (WPI) > 7 and
Symptom Severity
(SS) Scale score > 5 or
2. WPI = 3-6 and SS > 9
3. Symptoms present at similar level for > 3 months
4. No other disorder explaining the pain .

 Catastrophizing is an exaggerated negative attention to symptoms.
 Pain catastrophizing significantly predicts pain severity, chronic illness-related disability & emotional distress.
 High fatigue catastrophizing is associated with high fatigue severity in breast cancer patients and predicts post cancer treatment fatigue.
 Cognitive behavioral intervention focus on addressing maladaptive thinking improve fatigue severity and depression in chronic fatigue syndrome.

 No study has explored the role of catastrophizing in influencing possible biologic correlates of pain and fatigue in
FM.
 Little is known about the mechanisms that can explain both pain and fatigue symptoms experienced by FM patients.
 Few studies have looked at possible genomic correlates of
FM.




Questionnaires:
 Pain severity subscale of Brief Pain Inventory (BPI)
4-item subscale; the mean subscale score ranged from
0 to 10. The cutoff score that is clinically significant is 5.
 General fatigue subscale - Multidimensional Fatigue
Inventory (MFI) 4-item subscale; score range from 4-20.
The score of ≥ 13 is significant fatigue.
 Pain Catastrophizing Scale (PCS) = 13-item questionnaire; scores ranged from 0 - 52. Suggested clinical cut point is 16.

Biological sample collection and analysis:
 Microarray technology using
Affymetrix GeneChip ® human genome U133 Plus 2.0 array was used on blood collected using Paxgene ® tubes.
Whole blood
Collected using
Paxgene tubes
Microarray
 Differential gene expression was analyzed using Partek software.
 Gene selection criteria: over 2fold increase or decrease, p <
0.05.
 Network analyses by Ingenuity® software.
Extracted RNA
U133 Plus 2.0

9 Caucasian, female, diagnosed with FM, 26-64 years old
Min Max Mean (SD) Clinical
Cut-off
General Fatigue 13 20 17.1 (2.7) >13
Pain severity 0.5
6.3
4.1 (1.9)
Catastrophizing 4.0
36.0
17.0 (9.8)
>5
>16

High Pain Severity vs. Low Pain Severity (112 genes)
Up- regulated genes Down- regulated genes
Gene
Symbol
BATF2
Gene Title Function pvalue
Fold-
Change basic leucine zipper transcription factor, ATFlike 2
Protein binding/ protein dimerization activity
0.0002
2.1
CASP5 caspase 5, apoptosisrelated cysteine peptidase
Cyteine-type endopeptidase activity
0.0003
2.7
CCR1 chemokine
(C-C motif) receptor 1
CEACAM1 carcinoembry onic antigenrelated cell adhesion molecule 1
Molecular function, protein binding
0.001
2.4
COMMD6
COMM domain containing 6
C-C chemokine binding, chemokine (C-C motif) activity
0.0004
2.2
NF-kappa-B binding/ protein binding
0.001
4.1
Gene
Symbol
RPL7
SH2D1B
SIGLEC1 sialic acid binding Iglike lectin 1, sialoadhesin
UQCRB
ZCCHC2
Gene Title ribosomal protein L7
SH2 domain containing 1B ubiquinolcytochrome c reductase binding protein zinc finger,
CCHC domain containing 2
Function protein dimerization activity
Immune responses
Carbonydrate binding ubiquinolcytochrome-c reductase activity
Nucleic acid binding pvalue
Fold-
Change
0.02
0.04
0.04
0.05
0.05
-2.9
-2.8
-2.7
-2.6
-2.6

Network Analysis for high pain vs. low pain

High Catastrophizing vs. Low Catastrophizing (63 genes)
Up- regulated genes Down- regulated genes
Gene
Symbol
Gene Title function pvalue
Fold-
Change
SPP1 secreted phosphoprotein 1
Cytokine activity
0.01
2.1
TMTC1 transmembrane and
Integral to tetratricopeptide repeat containing membrane
1
0.02
2.5
SLC6A8 solute carrier family 6
(neurotransmitter transporter activity/ transporter,
Creatine molecular creatine), function member 8
0.03
2.4
NPRL3 nitrogen permease regulator-like 3
(S. cerevisiae)
Molecular function
0.04
2.1
HEMGN hemogen
Protein binding
0.04
2.2
Gene
Symbol
USP46
SEPT10
Gene Title ubiquitin specific peptidase 46 septin 10 function p-value
Fold-
Change
Ubiquitinspecific protease activity
GTP binding
0.001
0.002
-2.2
-2.0
CLEC4D
C-type lectin domain family 4, member D
Carbonydrate binding
0.005
-2.3
ZDHHC2 zinc finger,
DHHC-type containing 2
Zinc ion binding
0.005
-2.2
FAS
Fas (TNF receptor superfamily, member 6)
Identical protein binding/ signal transducer activity
0.010
-2.0

Network Analysis high vs. low catastrophizing

High vs Low Pain
 Basic leucine zipper protein
(BATF2) is regulated by interferon and serves as a suppressor of activating protein-1
(AP-1).
 Caspase 5, apoptosisrelated cysteine peptidase
(CASP5) and chemokine (C-C motif) receptor 1 (CCR1) are associated with immune response and inflammation in musculoskeletal disorders.
High vs Low Catastrophizing
 Secreted phosphoprotein 1
(SPP1 or Osteopontin) is upregulated during inflammation and associated with muscular dystrophies.
 Ubiquitin specific peptidase
46 (USP46) is a GABA regulation gene. In animal study, deletion of
USP46 is associated with depression-like behaviors in mice and rats.

STRESSOR
Inflammation
Inflammatory Cells: Leukocytes,
Lymphocytes, Platelets, Mast
Cells, Macrophages
CASP5
CCR1
BATF2
SPP1
Cytokines, Nerve Growth factor, prostaglandins,
Thromboxanes,
Leukotrienes, Serotonin
Hypothalamic-pituitary-adrenal (HPA) axis
Hypothalamus
Corticotropin releasing factors (CRF)
Anterior pituitary
Pituitary adrenocorticotropin
(ACTH)
Adrenal cortex
Physiological responses:
Pain and fatigue
USP46
Glucocorticoids
Fibromyalgia
Sympathetic nervous system
Adrenal gland:
Medulla
Acetylcholine (Ach) epinephrine norepinephrine
Bloodstream
Lymphoid organs
Behavioral responses:
Catastrophizing, depression

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


Leorey N. Saligan, PhD, CRNP
Tenure-Track Investigator
National Institute of Nursing Research, Intramural Research Program
Brian Walitt, MD, MPH, FACR
Associate Professor of Medicine
Georgetown University Medical Center
Benjamin Majors, BS
National Institute of Nursing Research, Intramural Research Program
Swarnalatha Reddy, PhD
National Institute of Nursing Research, Intramural Research Program

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