Gene expression profiling of liver from GSD1a patients: evidence of an inflammatory process

The world of

“omics”

International meeting Glycogen Storage Diseases associationsInternational meeting Glycogen Storage Diseases associationsMilan October 2Milan October 2ndnd 2010 2010

Luigi VaresioLuigi Varesio

54000 probsets54000 probsets

The chipThe chip

Gene expression Gene expression profile of GSD1a profile of GSD1a

liverliver

Gene expression profile of the liver of three patients who underwent liver transplantation and of three normal tissues.

Data analysis with Bibliosphere (Genomatics) to study metabolic pathways.

Gene expression profile of the liver of three patients who underwent liver transplantation and of three normal tissues.

Data analysis with Bibliosphere (Genomatics) to study metabolic pathways.

GSD1aClinical manifestations hepatomegaly

hypoglycemiahyperuricemiahyperlipidemiagrowth retardation

Complicationslack of growthdelay of pubertygout juvenilepancreatitisarteriosclerosishepatomashypertensionadenomaskidney diseases

G6PaseG6Pase

sugars

fat

proteins

??

What causes complications?

What causes complications?

0

5

10

15

20

25

30

35

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22IL6 Control

patient

IL-6

IL6 in GSD1a patients

IL6 Control

IL6 GSD1a

GSD1a patients have higher levels of IL-6GSD1a patients have higher levels of IL-6

Possible state of chronic inflammationPossible state of chronic inflammation

CX3CL1 chemokine (C-X3-C motif) ligand 1

CCL2 chemokine (C-C motif) ligand 2

CMKOR1 chemokine orphan receptor 1

CXCR4 chemokine (C-X-C motif) receptor 4

CXCR4 chemokine (C-X-C motif) receptor 4

XCL1 chemokine (C motif) ligand 1

CCL14 chemokine (C-C motif) ligand 15

CCL3 chemokine (C-C motif) ligand 3

CCL4 chemokine (C-C motif) ligand 4

CXCL9 chemokine (C-X-C motif) ligand 9

CRLF3 cytokine receptor-like factor 3

SOCS3 suppressor of cytokine signaling 3

CISH cytokine inducible SH2-containing protein

IL17RC interleukin 17 receptor C

IL1RAPL1 interleukin 1 receptor accessory protein-like 1

IL1RN interleukin 1 receptor antagonist

IL8Homo sapiens interleukin 8 C-terminal variant (IL8) mRNA, complete cds.

IL10 interleukin 10

IL18RAP interleukin 18 receptor accessory protein

IL8RB interleukin 8 receptor, beta

IL1R2 interleukin 1 receptor, type II

IL6 interleukin 6 (interferon, beta 2)

IL10RA interleukin 10 receptor, alpha

IL1RL1LG interleukin 1 receptor-like 1 ligand

IL8 interleukin 8

ILF2 interleukin enhancer binding factor 2, 45kDa

GSD1a can be associated with an inflammatory state

GSD1a can be associated with an inflammatory state

IL-6 transgenic mice

0

10

20

30

1 14 28 42 56

Age (day)

Bo

dy

wei

gh

t (g

r)

Wild-type Transgenic

De Benedetti, J Clin Invest 1997

12 day-old littermates

GSD1aClinical manifestations hepatomegaly

hypoglycemiahyperuricemiahyperlipidemiagrowth retardation

Complicationslack of gowthdelay of pubertygout juvenilepancreatitisarteriosclerosishepatomashypertensionadenomaskidney diseases

sugars

fats

proteins

G6PaseG6Pase

InflammationInflammation

CytokinesIL-6, IL-8

Hypoxia in patho-physiological processesHypoxia in patho-physiological processesHypoxia in patho-physiological processesHypoxia in patho-physiological processes

HYPOXIA(pO2 0-20mm Hg)

Growing tumors

Cerebral ischemia Myocardial ischemia

Pulmonary hypertension

Rheumatoid arthritis

Diabetic retinopathy

Wound healing

PreeclampsiaIntrauterine growth

retardation

Energy metabolism

Cell proliferation/survival

Angiogenesis Cell survival

Cell migrationErytropoiesis

Microbial infections

Aherosclerosis

Hypoxia in the liver of GSD1a patients Hypoxia in the liver of GSD1a patients Hypoxia in the liver of GSD1a patients Hypoxia in the liver of GSD1a patients

NAMENAME

23 GENI OXP23 GENI OXP

HYPOXIA_REG_UPHYPOXIA_REG_UP

LINES_MYC_LEALINES_MYC_LEA

NB-HYPO_32 GENESNB-HYPO_32 GENES

L1L2_11PBSTL1L2_11PBST

HYPOXIA_RCC_NOVHL_UPHYPOXIA_RCC_NOVHL_UP

L1L2_M-L1L2_M-

V$HIF1_Q3V$HIF1_Q3

HYPOXIA_NORMAL_UPHYPOXIA_NORMAL_UP

HIF1_TARGETSHIF1_TARGETS

L1L2_M+L1L2_M+

TRP_U133TRP_U133

HYPOXIA_REVIEWHYPOXIA_REVIEW

V$HIF1_Q5V$HIF1_Q5

HYPOXIA_RCC_UPHYPOXIA_RCC_UP

LEA_C1_SET_TOPLEA_C1_SET_TOP

NAME

HSA00190_OXIDATIVE_PHOSPHORYLATION

HSA00330_ARGININE_AND_PROLINE_METABOLISM

HSA01040_POLYUNSATURATED_FATTY_ACID_BIOSYNTHESIS

HSA00272_CYSTEINE_METABOLISM

HSA00360_PHENYLALANINE_METABOLISM

HSA00010_GLYCOLYSIS_AND_GLUCONEOGENESIS

HSA00020_CITRATE_CYCLE

HSA00910_NITROGEN_METABOLISM

HSA00252_ALANINE_AND_ASPARTATE_METABOLISM

HSA00030_PENTOSE_PHOSPHATE_PATHWAY

HSA04610_COMPLEMENT_AND_COAGULATION_CASCADES

HSA00220_UREA_CYCLE_AND_METABOLISM_OF_AMINO_GROUPS

OO22

CC-chemokine receptors ↓

Gro-chemokines ↑

Monocyte trapping

Circulation

Monocyte

Hypoxic tissue

Mn migration inhibiting factor (MIF) ↑G-protein deactivator (RGS1) ↑

Motility-promoting factor (ENPP2) ↓

CXC-Chemokines ↑

Neutrophils recruitment

Necrotic foci, elevated chemokines and infiltrating neutrophils in the

liver of glycogen storage disease type Ia.

Kim DY et al, J Hepatol. 2008Necrotic foci, elevated chemokines and infiltrating neutrophils in the

liver of glycogen storage disease type Ia.

Kim DY et al, J Hepatol. 2008

What causes these effects?

What causes these effects?

Liver hypoxia canLiver hypoxia can

GSD1aClinical manifestations hepatomegaly

hypoglycemiahyperuricemiahyperlipidemiagrowth retardation

Complicationslack of gowthdelay of pubertygout juvenilepancreatitisarteriosclerosishepatomashypertensionadenomaskidney diseases

sugars

fats

proteins

G6PaseG6Pase

InflammationInflammation

CytokinesIL-6, IL-8

Liver Liver HypoxiaHypoxia

IL-6

IL-6Rgp130

sIL-6R

MRA (Tocilizumab)a humanized moAb to the IL-6R

YY

YYYY

MRA

gp130

CollaboratorsCollaboratorsCollaboratorsCollaborators

Molecular Biology Gaslini Molecular Biology Gaslini • R. LavieriR. Lavieri• A. EvaA. Eva• P. FardinP. Fardin• R.ResazR.Resaz• L. VaresioL. Varesio

Molecular Biology Gaslini Molecular Biology Gaslini • R. LavieriR. Lavieri• A. EvaA. Eva• P. FardinP. Fardin• R.ResazR.Resaz• L. VaresioL. Varesio

NationalNational Institutes of Health Institutes of Health (USA)(USA)• J.ChouJ.Chou• D. MunroeD. Munroe• S.K. AryaS.K. Arya

NationalNational Institutes of Health Institutes of Health (USA)(USA)• J.ChouJ.Chou• D. MunroeD. Munroe• S.K. AryaS.K. Arya

IST Genova IST Genova • O. BarbieriO. Barbieri

IST Genova IST Genova • O. BarbieriO. Barbieri

Clinical DepartmentsClinical Departments

Dr. M. Di RoccoDr. M. Di Rocco• G.Gaslini Institute (Genova)G.Gaslini Institute (Genova)

Dr. D. MelisDr. D. Melis

• Federico II University (Napoli) Federico II University (Napoli)

Dr. R.PariniDr. R.Parini

•S.Gerardo Hospital (Monza)S.Gerardo Hospital (Monza)

Dr. S. PaciDr. S. Paci

•S.Paolo Hospital (Milano)S.Paolo Hospital (Milano)

Dr. E. ProcopioDr. E. Procopio

•Meyer Hospital (Firenze)Meyer Hospital (Firenze)

Dr. C. DionisiDr. C. Dionisi

•Bambin Gesù Hospital (Roma)Bambin Gesù Hospital (Roma)

Clinical DepartmentsClinical Departments

Dr. M. Di RoccoDr. M. Di Rocco• G.Gaslini Institute (Genova)G.Gaslini Institute (Genova)

Dr. D. MelisDr. D. Melis

• Federico II University (Napoli) Federico II University (Napoli)

Dr. R.PariniDr. R.Parini

•S.Gerardo Hospital (Monza)S.Gerardo Hospital (Monza)

Dr. S. PaciDr. S. Paci

•S.Paolo Hospital (Milano)S.Paolo Hospital (Milano)

Dr. E. ProcopioDr. E. Procopio

•Meyer Hospital (Firenze)Meyer Hospital (Firenze)

Dr. C. DionisiDr. C. Dionisi

•Bambin Gesù Hospital (Roma)Bambin Gesù Hospital (Roma)

Associazione Associazione Italiana Italiana

GlicogenosiGlicogenosi