AKO ĎALEKO JE K APLIKOVANÉMU
VÝSKUMU
MN/CA IX story
Jaromír Pastorek
IDENTIFICATION OF CARBONIC ANHYDRASE IX
CA IX (MN)
Transmembrane 58/54 kDa glycoprotein, induced by HeLa cell density,
trimers in non-reducing conditions
Pastoreková, Závadová, Košťál, Babušíková,
Závada: Virology 187, 620-6, 1992.
Y
M75
Expression correlates with the tumorigenic phenotype
and is associated with tumor tissues
Závada, Závadová, Pastoreková, Čiampor,
Pastorek, Zelník: Int J Cancer 54, 268-274, 1993.
Liao, Brewer, Zavada, Pastorek,
Pastorekova, Manetta, Berman,
DiSaia, Stanbridge:
Am J Pathol 145, 598-609, 1994.
Potential diagnostic utility in early detection of cervical carcinoma
Primary structure of cDNA and gene organization → protein composition → CA IX
Pastorek, Pastorekova, Callebaut, Mornon, Zelnik, Opavsky, Zat'ovicova, Liao, Portetelle, Stanbridge, Zavada, Burny, Kettmann:
Oncogene 9, 2877-88, 1994.
Opavsky*, Pastorekova*, Zelnik, Gibadulinova, Stanbridge, Zavada, Kettmann, Pastorek: Genomics 33, 480-487.
signal peptide
proteoglycan-like region - PG
carbonic anhydrase domain - CA
CO2 + H2O ⇔ HCO3- + H+
transmembrane anchor - TM
intracytoplasmic tail - IC
HUMAN CARBONIC ANHYDRASES
M75
DISTRIBUTION OF CA IX IN HUMAN TISSUES
Pastorekova, Parkkila,
Parkkila, Opavsky,
Zelnik, Saarnio, Pastorek:
Gastroenterology 112,
398-408, 1997
Saarnio, Parkkila, Parkkila,
Waheed, Casey, Zhou,
Pastorekova, Pastorek,
Karttunen, Haukipuro,
Kairaluoma, Sly:
J Histochem Cytochem 46,
497-504, 1998
Bartosova, Parkkila,
Pohlodek, Karttunen,
Galbavy, Mucha,
Harris, Pastorek,
Pastorekova:
J Pathol 197, 1-8, 2002.
Saarnio, Parkkila, Parkkila,
Haukipuro, Pastorekova,
Pastorek, Kairaluoma,
Karttunen: Am J Pathol
153, 279-285, 1998.
REGULATION OF CA IX EXPRESSION - TRANSCRIPTION
Kaluz, Kaluzova, Opavsky, Pastorekova, Gibadulinova, Dequiedt, Kettmann,
Pastorek: J Biol Chem 274, 32588-32595, 1999.
Wykoff, Beasley, Watson, Turner, Pastorek, Wilson, Turley, Maxwell, Pugh,
Ratcliffe, Harris A: Cancer Res 60, 7075-7083.
Kaluz 2002
Kopacek 2005
hypoxia
(non-RCC)
MAPK
pVHL (RCC)
PI3K
HIF-1
cell density
α
mRNA
methylation (RCC)
SP1
HIF-1
β α
REGULATION OF CA IX EXPRESSION - SPLICING
Barathova et al, BJC 2007, in press
REGULATION OF CA IX EXPRESSION – STABILITY & SHEDDING
Rafajova, Zatovicova et al, IJO 2004, in press
Zatovicova et al, BJC 2005
REGULATION OF CA IX EXPRESSION
Normoxia - splicing variant AS CA IX
Hypoxia, cell density - WT CA IX
Key transcription factors HIF & SP1
Post-translation – shedding
- phosphorylation
- modulation of enzyme
activity
WT CA IX
enzymaticly active in hypoxia
inactive in normoxia
AS CA IX
inactive
reduces activity of WT when
added to hypoxic cells
ROLE OF CA IX IN CELL ADHESION
neo
CA IX
Zaťovičová, Tarábková, Švastová, Gibadulinová, Jakubíčková, Biesová,
Rafajová, Mucha, Gut, Parkkila,
Waheed, Sly, Horak, Pastorek, Pastorekova: J Immunol Methods, 2003,
in press.
HYPOXIA
Cellular changes
Reduced proliferation
Anaerobic metabolism
Acidic microenvironment
Reduced cell adhesion
Angiogenesis
Cell death
Selection of aggressive
tumour cells
poor prognosis
Clinical consequences
worse responses to conventional
treatment modalities
Resistance to radiotherapy
and chemotherapy
blood vessels
doxorubicin
hypoxia
HIF-mediated molecular responses to hypoxia
Metabolic shift from oxidative phosphorylation
to anaerobic glycolysis
Purpose:
To maintain production
of energy
Consequence:
High production of
lactic acid, CO2 & protons
Glycolysis-deficient cells
Acidic tumors in vivo
pH regulation
Monocarboxylate
transporter
H+
Vacuolar
proton
pump
H+
H+
CO2
Na+/H+
antiport
H+
Bicarbonate
transporter
Purpose:
neutralize pHi
& survive
acidify pHe
& invade
and metastasize
CA IX is a component of bicarbonate metabolon
CA IX activity = inhibited by bicarbonate but not lactate
CO2 + H2O
HCO3- + H+
ROLE OF CA IX IN pH REGULATION
A
N
B
H
N
H
lactate
CA IX
mock
CA IX
MDCK
Normoxia
Hypoxia
mock
CA IX
C
Svastova et al-FEBS Let-2004
Hypoxia regulates
CA IX level
+
catalytic activity
CA IX–selective sulfonamides as promising tools
for imaging of hypoxic tumors in vivo
HeLa transfected with delCA variant
unpublished
5 min
IMMUNOTARGETING
1h
4°C
1
VII/20 MAb
CA IX
2
37°C
r r ecy
p to
ng
c li
rec
e
20 min
P
3
?
io
n
degrad a
37°C
+
0.2 M
acetic acid
ly endoso
e
li ga
nd
ar
m
e
6
t
4
low pH
5
l ysos ome
ligan d
a
nd
pt or d
e
cytoplasm
control
MAb IV/18 MAb V/10 MAb VII/20
1
fl CA IX
∆CA
2
1
2
1
2
Tumor volume
nucleus
rece
gr
ad
a tion
e
nd
osome
MAb VII/20
(10 mg/dose)
3h
CA IX as a screening/monitoring marker
Li G, Feng G, Gentil-Perret A, Genin C, Tostain J.
J Urol. 2008 Aug;180(2):510-3
1,2
1
Mean serum levels:
Metastatic RCC patients 216.68 +/- 67.02 pg/ml or
Localized RCC patients 91.65 +/- 13.29 pg/ml
Healthy individuals
14.59 +/- 6.22 pg/ml
p <0.001 and p = 0.001, respectively
0,8
0,6
0,4
Higher levels = higher recurrence
0,2
0
1
tumor
3
5
7
9
11
shed ECD
13
15
17
19
21
3,5
3,5
3
3
2,5
2,5
2
2
1,5
1,5
1
1
0,5
0,5
Dilutions 1:101:10-320
No.15 CA IX
No.16 neo
0
0
1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16
1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16
DNEŠNÝ STATUS CAIX
- najlepší marker nádorovej hypoxie
- marker zlej prognózy (agresívne nádory)
- terč pre cielenú liečbu nádorov
VÝSLEDKY a PUBLIKÁCIE
PROJEKTY a PÍSANIE
PATENTY a FIRMY a PAPIEROVANIE atď.
Základný výskum – granty VEGA
APVV
6. RP EU Euroxy
7. RP EU Metoxia
medzinárodná spolupráca:
Belgicko, Fínsko, Taliansko, UK....
Výsledok - publikácie
citácie
medzinárodná spolupráca
získavanie projektov
patentový podklad
Pastoreková S, Závadová Z, Kostál M, Babusíková O, Závada J. A novel quasi-viral agent, MaTu, is a two-component system. Virology. 1992
Apr;187(2):620-6.
Zavada J, Zavadova Z, Pastorekova S, Ciampor F, Pastorek J, Zelnik V (1993). Expression of MaTu-MN protein in human tumor cultures and in
clinical specimens. Int J Cancer 54, 268-274.
Liao SY, Brewer C, Zavada J, Pastorek J, Pastorekova S, Manetta A, Berman ML, DiSaia PJ, Stanbridge EJ (1994). Identification of the MN
antigen as a diagnostic biomarker of cervical intraepithelial squamous and glandular neoplasia and cervical carcinomas. Am J Pathol 145,
598-609.
Pastorek J, Pastorekova S, Callebaut I, Mornon JP, Zelnik V, Opavsky R, Zatovicova M, Liao S, Portetelle D, Stanbridge EJ, Zavada J, Burny A,
Kettmann R (1994). Cloning and characterization of MN, a human tumor-associated protein with a domain homologous to carbonic
anhydrase and a putative helix-loop-helix DNA binding segment. Oncogene 9, 2877-88.
Opavsky R, Pastorekova S, Zelnik V, Gibadulinova A, Stanbridge EJ, Zavada J, Kettmann R, Pastorek J (1996). Human MN/CA9 gene, a novel
member of the carbonic anhydrase family: structure and exon to protein domain relationships. Genomics 33, 480-487.
Brewer CA, Liao SY, Wilczynski SP, Pastorekova S, Pastorek J, Zavada J, Kurosaki T, Manetta A, Berman ML, DiSaia PJ, Stanbridge EJ (1996).
A study of biomarkers in cervical carcinoma and clinical correlation of the novel biomarker MN. Gynecol Oncol 63, 337-44.
Pastorekova S, Parkkila S, Parkkila AK, Opavsky R, Zelnik V, Saarnio J, Pastorek J (1997). Carbonic anhydrase IX, MN/CA IX: analysis of
stomach complementary DNA sequence and expression in human and rat alimentary tracts. Gastroenterology 112, 398-408.
Saarnio J, Parkkila S, Parkkila AK, Waheed A, Casey MC, Zhou XY, Pastorekova S, Pastorek J, Karttunen T, Haukipuro K, Kairaluoma MI, Sly
WS (1998). Immunohistochemistry of carbonic anhydrase isozyme IX (MN/CA IX) in human gut reveals polarized expression in the epithelial
cells with the highest proliferative capacity. J Histochem Cytochem 46, 497-504.
Saarnio J, Parkkila S, Parkkila AK, Haukipuro K, Pastorekova S, Pastorek J, Kairaluoma MI, Karttunen TJ (1998). Immunohistochemical study of
colorectal tumors for expression of a novel transmembrane carbonic anhydrase, MN/CA IX, with potential value as a marker of cell
proliferation. Am J Pathol 153, 279-285.
Lieskovska J, Kaluzova M, Opavsky R, Kaluz S, Pastorek J, Kettmann R, Pastorekova S (1998). Up-regulation of p53 by antisense expression of
HPV18 E6 oncogene does not influence the level of MN/CA IX tumor-associated protein in HeLa cervical carcinoma cells. Int J Oncol 13,
1081-1086.
Kaluz S, Kaluzová M, Pastorek J. Inverse PCR-generated internally deleted constructs for direct characterization of promoter regulatory regions.
Biotechniques. 1999 Mar;26(3):446-50.
Lieskovska J, Opavsky R, Zacikova L, Glasova M, Pastorek J, Pastorekova S (1999). Study of in vitro conditions modulating expression of MN/CA
IX protein in human cell lines derived from cervical carcinoma. Neoplasma 46, 17-24.
Kaluz S, Kaluzova M, Opavsky R, Pastorekova S, Gibadulinova A, Dequiedt F, Kettmann R, Pastorek J (1999). Transcriptional regulation of the
MN/CA 9 gene coding for the tumor-associated carbonic anhydrase IX. Identification and characterization of a proximal silencer element. J
Biol Chem 274, 32588-32595.
Parkkila S, Rajaniemi H, Parkkila AK, Kivela J, Waheed A, Pastorekova S, Pastorek J, Sly WS (2000). Carbonic anhydrase inhibitor suppresses
invasion of renal cancer cells in vitro. Proc Natl Acad Sci U S A. 97, 2220-2224.
Wykoff CC, Beasley NJ, Watson PH, Turner KJ, Pastorek J, Sibtain A, Wilson GD, Turley H, Talks KL, Maxwell PH, Pugh CW, Ratcliffe PJ, Harris
AL. Hypoxia-inducible expression of tumor-associated carbonic anhydrases. Cancer Res. 2000 Dec 15;60(24):7075-83.
Závada J, Závadová Z, Pastorek J, Biesová Z, Jezek J, Velek J. Human tumour-associated cell adhesion protein MN/CA IX: identification of M75
epitope and of the region mediating cell adhesion. Br J Cancer. 2000 Jun;82(11):1808-13.
Kaluzova M, Pastorekova S, Pastorek J, Kaluz S (2000). P53 tumour suppressor modulates transcription of the TATA-less gene coding for the
tumour-associated carbonic anhydrase MN/CA IX in MaTu cells. Biochim Biophys Acta 1491, 20-26.
Kivela AJ, Parkkila S, Saarnio J, Karttunen TJ, Kivela J, Parkkila AK, Pastorekova S, Pastorek J, Waheed A, Sly WS, Rajaniemi H (2000).
Expression of transmembrane carbonic anhydrase
isoenzymes IX and XII in normal human pancreas and pancreatic tumours. Histochem Cell Biol 114, 197-204.
Wykoff CC, Beasley N, Watson PH, Campo L, Chia SK, English R, Pastorek J, Sly WS, Ratcliffe P, Harris AL. Expression of the hypoxia-inducible and tumorassociated carbonic anhydrases in ductal carcinoma in situ of the breast. Am J Pathol. 2001 Mar;158(3):1011-9.
Karhumaa P, Kaunisto K, Parkkila S, Waheed A, Pastorekova S, Pastorek J, Sly WS, Rajaniemi H (2001). Expression of the transmembrane carbonic anhydrases,
CA IX and CA XII, in the human male excurrent ducts. Mol Hum Reprod 7, 611-616.
Beasley NJ, Wykoff CC, Watson PH, Leek R, Turley H, Gatter K, Pastorek J, Cox GJ, Ratcliffe P, Harris AL. Carbonic anhydrase IX, an endogenous hypoxia marker,
expression in head and neck squamous cell carcinoma and its relationship to hypoxia, necrosis, and microvessel density. Cancer Res. 2001 Jul 1;61(13):52627.
Chia SK, Wykoff CC, Watson PH, Han C, Leek RD, Pastorek J, Gatter KC, Ratcliffe P, Harris AL. Prognostic significance of a novel hypoxia-regulated marker,
carbonic anhydrase IX, in invasive breast carcinoma. J Clin Oncol. 2001 Aug 15;19(16):3660-8.
Loncaster JA, Harris AL, Davidson SE, Logue JP, Hunter RD, Wycoff CC, Pastorek J, Ratcliffe PJ, Stratford IJ, West CM. Carbonic anhydrase (CA IX) expression, a
potential new intrinsic marker of hypoxia: correlations with tumor oxygen measurements and prognosis in locally advanced carcinoma of the cervix. Cancer
Res. 2001 Sep 1;61(17):6394-9.
Kaluzova M, Pastorekova S, Svastova E, Pastorek J, Stanbridge EJ, Kaluz S (2001). Characterization of the MN/CA 9 promoter proximal region: a role for specificity
protein (SP) and activator protein 1 (AP1) factors. Biochem J 359, 669-677.
Kivela AJ, Saarnio J, Karttunen TJ, Kivela J, Parkkila AK, Pastorekova S, Pastorek J, Waheed A, Sly WS, Parkkila TS, Rajaniemi H (2001). Differential expression of
cytoplasmic carbonic anhydrases, CA I and II, and membrane-associated isozymes, CA IX and XII, in normal mucosa of large intestine and in colorectal
tumors. Dig Dis Sci 46, 2179-2186.
Saarnio J, Parkkila S, Parkkila AK, Pastorekova S, Haukipuro K, Pastorek J, Juvonen T, Karttunen TJ (2001).Transmembrane carbonic anhydrase, MN/CA IX, is a
potential biomarker for biliary tumours. J Hepatol 35, 643-649.
Giatromanolaki A, Koukourakis MI, Sivridis E, Pastorek J, Wykoff CC, Gatter KC, Harris AL. Expression of hypoxia-inducible carbonic anhydrase-9 relates to
angiogenic pathways and independently to poor outcome in non-small cell lung cancer. Cancer Res. 2001, 61(21):7992-8.
Koukourakis MI, Giatromanolaki A, Sivridis E, Simopoulos K, Pastorek J, Wykoff CC, Gatter KC, Harris AL. Hypoxia-regulated carbonic anhydrase-9 (CA9) relates to
poor vascularization and resistance of squamous cell head and neck cancer to chemoradiotherapy. Clin Cancer Res. 2001 Nov;7(11):3399-403.
Turner KJ, Crew JP, Wykoff CC, Watson PH, Poulsom R, Pastorek J, Ratcliffe PJ, Cranston D, Harris AL. The hypoxia-inducible genes VEGF and CA9 are
differentially regulated in superficial vs invasive bladder cancer. Br J Cancer. 2002 Apr 22;86(8):1276-82.
Leppilampi M, Koistinen P, Savolainen ER, Hannuksela J, Parkkila AK, Niemela O, Pastorekova S, Pastorek J, Waheed A, Sly WS, Rajaniemi H, Parkkila S (2002).
Carbonic anhydrase in hematologic malignancies. Clin Canc Res 8, 2240-2245.
Bartosova M, Parkkila S, Pohlodek K, Karttunen TJ, Galbavy S, Mucha V, Harris A, Pastorek J, Pastorekova S (2002). Expression of carbonic anhydrase IX in breast
is associated with malignant tissues and is related to overexpression of c-erbB2. J Pathol 197, 1-8.
Kaluz S, Kaluzova M, Chrastina A, Olive PL, Pastorekova S, Pastorek J, Lerman MI, and Stanbridge EJ (2002) Lowered oxygen tension induces expression of the
hypoxia marker MN/carbonic anhydrase IX in the absence of hypoxia-inducible factor 1α stabilization : a role for phosphatidylinositol 3’-kinase. Cancer Res
62, 4469-4477.
Ortova Gut M, Parkkila S, Vernerova Z, Rohde E, Závada J, Höcker M, Pastorek J, Karttunen T, Zavadova Z, Knobeloch KP, Wiedenmann B, Svoboda J, Horak I,
and Pastorekova S (2002). Gastric hyperplasia in mice with targeted disruption of the carbonic anhydrase gene Car9. Gastroenterology 123, 1889-1903.
Swinson DE, Jones JL, Richardson D, Wykoff C, Turley H, Pastorek J, Taub N, Harris AL, O'Byrne KJ. Carbonic anhydrase IX expression, a novel surrogate marker
of tumor hypoxia, is associated with a poor prognosis in non-small-cell lung cancer. J Clin Oncol. 2003 Feb 1;21(3):473-82.
Vullo D, Franchi M, Gallori E, Pastorek J, Scozzafava A, Pastorekova S, and Supuran C (2003) Carbonic anhydrase inhibitors. Inhibition of the tumorassociated isozyme IX with aromatic and heterocyclic sulfonamides. Bioorg Med Chem Let 13, 1005-1009.
Ilies MA, Vullo D, Pastorek J, Scozzafava A, Ilies M, Caproiu T, Pastorekova S, and Supuran C (2003) Carbonic anhydrase inhibitors. Inhibition of tumorassociated isozyme IX by halogeno-sulfanilamide and halogeno-aminobenzolamide derivatives. J Med Chem, 46, 2187-96.
Chrastina A, Zavada J, Parkkila S, Kaluz S, Kaluzova M, Rajcani J, Pastorek J, and Pastorekova S (2003) Biodistribution and pharmacokinetics of 125Ilabeled monoclonal antibody M75 specific for carbonic anhydrase IX, an intrinsic marker of hypoxia, in nude mice xenografted with human
colorectal carcinoma. Int J Cancer 105, 873-881.
Zatovicova M, Tarabkova K, Svastova E, Gibadulinova A, Jakubickova L, Biesova Z, Rafajova M, Mucha V, Ortova Gut M, Horak I, Pastorek J,
Pastorekova S (2003) Monoclonal antibodies generated in CA IX-deficient mice recognize different domains of tumor-associated hypoxia-induced
carbonic anhydrase IX. J Immunol Methods, 282, 117-134.
Leppilampi M, Saarnio J, Karttunen TJ, Kivela J, Pastorekova S, Pastorek J, Waheed A, Sly WS, Parkkila S (2003) Carbonic anhydrase isozymes IX and
XII in gastric tumors. World J Gastroenterol 9,1398-1403.
Tomes L, Emberley E, Niu Y, Troup S, Pastorek J, Strange K, Harris A, Watson PH. Necrosis and hypoxia in invasive breast carcinoma. Breast Cancer
Res Treat. 2003 Sep;81(1):61-9.
Franchi M, Vullo D, Gallori E, Pastorek J, Russo A, Scozzafava A, Pastorekova S, Supuran CT (2003). Carbonic anhydrase inhibitors. Inhibition of
cytosolic isozymes I and II and transmembrane, cancer-associated isozyme IX with lipophilic sulfonamides. J Enz Inhib 18, 333-338.
Vullo D, Franchi M, Gallori E, Pastorek J, Russo A, Scozzafava A, Pastorekova S, Supuran CT (2003). Carbonic anhydrase inhibitors. Inhibition of
cytosolic isozymes I and II and transmemebrane, cancer-associated isozyme IX with anions. J Enz Inhib, 18, 403-406.
Chrastina A, Pastorekova S and Pastorek J (2003). Immunotargeting of human cervical carcinoma xenograft expressing CA IX tumor-associated antigen
by 125I-labeled M75 monoclonal antibody. Neoplasma 50, 13-21.
Svastová E, Zilka N, Zatovicova M, Gibadulinova A, Ciampor F, Pastorek J, Pastorekova S (2003). Carbonic anhydrase IX reduces E-cadherin-mediated
adhesion of MDCK cells via interaction with β-catenin. Exp Cell Res 290, 332-345.
Rafajova M, Zatovicova M, Kettmann R, Pastorek J, Pastorekova S (2004). Induction by hypoxia combined with low glucose or low bicarbonate and high
posttranslational stability upon reoxygenation contribute to carbonic anhydrase IX expression in cancer cells. Int J Oncol, 24, 995-1004.
Pastorekova S, Casini A, Scozzafava A, Vullo D, Pastorek J, Supuran CT (2004). Carbonic anhydrase inhibitors: The first selective, membraneimpermeant inhibitors targeting the tumor-associated isozyme IX. Bioorg Med Chem Lett 14, 869–873.
Hussain SA, Palmer DH, Ganesan R, Hiller L, Gregory J, Murray PG, Pastorek J, Young L, James ND. Carbonic anhydrase IX, a marker of hypoxia:
correlation with clinical outcome in transitional cell carcinoma of the bladder. Oncol Rep. 2004 May;11(5):1005-10.
Koukourakis MI, Giatromanolaki A, Sivridis E, Pastorek J, Karapantzos I, Gatter KC, Harris AL; Tumour and Angiogenesis Research Group. Hypoxiaactivated tumor pathways of angiogenesis and pH regulation independent of anemia in head-and-neck cancer. Int J Radiat Oncol Biol Phys. 2004
May 1;59(1):67-71.
Vullo D, Scozzafava A, Pastorekova S, Pastorek J, Supuran CT (2004). Carbonic anhydrase inhibitors: inhibition of the tumor-associated isozyme IX with
fluorine-containing sulfonamides. The first subnanomolar CA IX inhibitor discovered. Bioorg Med Chem Lett 14, 2351-6.
Hilvo M, Rafajova M, Pastorekova S, Pastorek J, Parkkila S (2004). Expression of carbonic anhydrase IX in mouse tissues. J Histochem Cytochem
52,1313-22.
Pohlodek K, Galbavy S, Bartosova M, Mucha V, Holoman K, Pastorek J, Pastorekova S (2004). Semi-quantitative RT-PCR assessment of molecular
markers in breast large-core needle biopsies. Neoplasma 51, 415-21.
Hynninen P, Hamalainen JM, Pastorekova S, Pastorek J, Waheed A, Sly WS, Tomas E, Kirkinen P, Parkkila S (2004). Transmembrane carbonic
anhydrase isozymes IX and XII in the female mouse reproductive organs. Reprod Biol Endocrinol 17,2(1):73.
Pastorekova S, Parkkila S, Pastorek J, Supuran CT (2004). Carbonic anhydrase: current state of the art, therapeutic applications and future prospects. J
Enz Inhib Med Chem, 19, 199-229. Review.
Svastova E, Hulikova A, Rafajova M, Zat'ovicova M, Gibadulinova A, Casini A, Cecchi A, Scozzafava A, Supuran CT, Pastorek J, Pastorekova S (2004).
Hypoxia activates the capacity of tumor-associated carbonic anhydrase IX to acidify extracellular pH. FEBS Lett 577, 439-45.
Winum JY, Pastorekova S, Jakubickova L, Montero JL, Scozzafava A, Pastorek J, Vullo D, Innocenti A, Supuran CT (2005). Carbonic anhydrase inhibitors:
synthesis and inhibition of cytosolic/tumor-associated carbonic anhydrase isozymes I, II, and IX with bis-sulfamates. Bioorg Med Chem Lett 15, 57984.
Vullo D, Innocenti A, Nishimori I, Pastorek J, Scozzafava A, Pastorekova S, Supuran CT (2005). Carbonic anhydrase inhibitors. Inhibition of the
transmembrane isozyme XII with sulfonamides-a new target for the design of antitumor and antiglaucoma drugs? Bioorg Med Chem Lett 15, 963-9.
Jakubickova L, Biesova Z, Pastorekova S, Kettmann R, Pastorek J (2005). Methylation of the CA9 promoter can modulate expression of the tumorassociated carbonic anhydrase IX in dense carcinoma cell lines. Int J Oncol 26, 1121-7.
Kopacek J, Barathova M, Dequiedt F, Sepelakova J, Kettmann R, Pastorek J, Pastorekova S (2005). MAPK pathway contributes to density- and hypoxiainduced expression of the tumor-associated carbonic anhydrase IX. Biochim Biophys Acta 1729, 41-9.
Kivela AJ, Parkkila S, Saarnio J, Karttunen TJ, Kivela J, Parkkila AK, Bartosova M, Mucha V, Novak M, Waheed A, Sly WS, Rajaniemi H, Pastorekova S,
Pastorek J (2005). Expression of von Hippel-Lindau tumor suppressor and tumor-associated carbonic anhydrases IX and XII in normal and
neoplastic colorectal mucosa. World J Gastroenterol 11, 2616-25.
Kowalewska M, Radziszewski J, Kulik J, Barathova M, Nasierowska-Guttmajer A,Bidzinski M, Pastorek J, Pastorekova S, Siedlecki JA (2005). Detection of
carbonic anhydrase 9-expressing tumor cells in the lymph nodes of vulvar carcinoma patients by RT-PCR. Int J Cancer 116, 957-62.
Cecchi A, Hulikova A, Pastorek J, Pastorekova S, Scozzafava A, Winum JY, Montero JL, Supuran CT (2005). Carbonic anhydrase inhibitors. Design of
fluorescent sulfonamides as probes of tumor-associated carbonic anhydrase IX that inhibit isozyme IX-mediated acidification of hypoxic tumors. J
Med Chem 48, 4834-41.
Pastorekova S, Vullo D, Casini A, Scozzafava A, Pastorek J, Nishimori I, Supuran CT (2005). Carbonic anhydrase inhibitors: Inhibition of the tumorassociated isozymes IX and XII with polyfluorinated aromatic/heterocyclic sulfonamides. J Enzyme Inhib Med Chem 20, 211-217.
Leppilampi M, Karttunen TJ, Kivela J, Gut MO, Pastorekova S, Pastorek J, Parkkila S (2005). Gastric pit cell hyperplasia and glandular atrophy in carbonic
anhydrase IX knockout mice: studies on two strains C57/BL6 and BALB/C. Transgenic Res 14, 655-63.
Zatovicova M, Sedlakova O, Svastova E, Ohradanova A, Ciampor F, Arribas J, Pastorek J, Pastorekova S (2005) Ectodomain shedding of the hypoxiainduced carbonic anhydrase IX is a metalloprotease-dependent process regulated by TACE/ADAM17. Br J Cancer 93, 1267-76.
Dorai T, Sawczuk IS, Pastorek J, Wiernik PH, Dutcher JP. The role of carbonic anhydrase IX overexpression in kidney cancer. Eur J Cancer. 2005
Dec;41(18):2935-47.
Pan P, Leppilampi M, Pastorekova S, Pastorek J, Waheed A, Sly WS, Parkkila S (2006). Carbonic anhydrase gene expression in CA II-deficient (Car2-/-)
and CA IX-deficient (Car9-/-) mice. J Physiol 571, 319-27.
Haapasalo JA, Nordfors KM, Hilvo M, Rantala IJ, Soini Y, Parkkila AK, Pastorekova S, Pastorek J, Parkkila SM, Haapasalo HK (2006). Expression of
carbonic anhydrase IX in astrocytic tumors predicts poor prognosis. Clin Cancer Res 15, 473-7.
Kallio H, Pastorekova S, Pastorek J, Waheed A, Sly WS, Mannisto S, Heikinheimo M, Parkkila S (2006). Expression of carbonic anhydrases IX and XII
during mouse embryonic development. BMC Dev Biol. 23, 6, 22.
Dorai T, Sawczuk I, Pastorek J, Wiernik PH, Dutcher JP. Role of carbonic anhydrases in the progression of renal cell carcinoma subtypes: proposal of a
unified hypothesis. Cancer Invest. 2006 Dec;24(8):754-79. Review.
Ihnatko R, Kubes M, Takacova M, Sedlakova O, Sedlak J, Pastorek J, Kopacek J, Pastorekova S (2006). Extracellular acidosis elevates carbonic
anhydrase IX in human glioblastoma cells via transcriptional modulation that does not depend on hypoxia. Int J Oncol. 29, 1025-33.
Hynninen P, Vaskivuo L, Saarnio J, Haapasalo H, Kivelä J, Pastorekova S, Pastorek J, Waheed A, Sly WS, Puistola U, Parkkila S (2006). Expression of
transmembrane carbonic anhydrases IX and XII in ovarian tumours. Histopathology 49, 594-602.
Labudova M, Tomaskova J, Kaluzova M, Pastorek J, Pastorekova S (2006). Lymphocytic choriomeningitis virus MX strain does not induce the expression
of tumor-associated carbonic anhydrase IX in persistently infected HeLa cells. Acta Virol 50, 53-8.
Hussain SA, Ganesan R, Reynolds G, Gross L, Stevens A, Pastorek J, Murray PG, Perunovic B, Anwar MS, Billingham L, James ND, Spooner D, Poole CJ,
Rea DW, Palmer DH. Hypoxia-regulated carbonic anhydrase IX expression is associated with poor survival in patients with invasive breast cancer. Br
J Cancer. 2007 Jan 15;96(1):104-9.
Innocenti A, Vullo D, Pastorek J, Scozzafava A, Pastorekova S, Nishimori I, Supuran CT (2007). Carbonic anhydrase inhibitors. Inhibition of transmembrane
isozymes XII (cancer-associated) and XIV with anions. Bioorg Med Chem Lett. 17, 1532-7.
Pastorekova S, Kopacek J, Pastorek J (2007). Carbonic anhydrase inhibitors and the management of cancer. Curr Top Med Chem. 7, 865-78. Review.
Niemelä AM, Hynninen P, Mecklin JP, Kuopio T, Kokko A, Aaltonen L, Parkkila AK, Pastorekova S, Pastorek J, Waheed A, Sly WS, Orntoft TF, Kruhøffer M,
Haapasalo H, Parkkila S, Kivelä AJ (2007). Carbonic anhydrase IX is highly expressed in hereditary nonpolyposis colorectal cancer. Cancer Epidemiol
Biomarkers Prev. 16, 1760-6.
Takacova M, Barathova M, Hulikova A, Ohradanova A, Kopacek J, Parkkila S, Pastorek J, Pastorekova S, Zatovicova M (2007). Hypoxia-inducible
expression of the mouse carbonic anhydrase IX demonstrated by new monoclonal antibodies. Int J Oncol. 31, 1103-10.
Barathova M, Takacova M, Holotnakova T, Gibadulinova A, Ohradanova A, Zatovicova M, Hulikova A, Kopacek J, Parkkila S, Supuran CT, Pastorekova S,
Pastorek J (2008). Alternative splicing variant of the hypoxia marker carbonic anhydrase IX expressed independently of hypoxia and tumour
phenotype. Br J Cancer. 98, 129-36.
Holotnakova T, Ziegelhoffer A, Ohradanova A, Hulikova A, Novakova M, Kopacek J, Pastorek J
J, Pastorekova S (2008). Induction of carbonic anhydrase IX
by hypoxia and chemical disruption of oxygen sensing in rat fibroblasts and cardiomyocytes. Pflugers Arch. 456, 323-37
Jarvela S, Parkkila S, Bragge H, Kahkonen M, Parkkila AK, Soini Y, Pastorekova S, Pastorek J, Haapasalo H (2008). Carbonic anhydrase IX in
oligodendroglial brain tumors. BMC Cancer. 8(1):1
Pastorekova S, Vullo D, Nishimori I, Scozzafava A, Pastorek J, Supuran CT. (2008) Carbonic anhydrase activators: activation of the human tumorassociated isozymes IX and XII with amino acids and amines. Bioorg Med Chem. 16(7):3530-6.
Haapasalo J, Hilvo M, Nordfors K, Haapasalo H, Parkkila S, Hyrskyluoto A, Rantala I, Waheed A, Sly WS, Pastorekova S, Pastorek J, Parkkila AK. (2008)
Identification of an alternatively spliced isoform of carbonic anhydrase XII in diffusely infiltrating astrocytic gliomas. Neuro Oncol. 10(2):131-8.
Pastorekova S, Ratcliffe PJ, Pastorek J.(2008) Molecular mechanisms of carbonic anhydrase IX-mediated pH regulation under hypoxia. BJU Int. 101 Suppl
4:8-15.
Pastorekova S, Zatovicova M, Pastorek J. (2008) Cancer-associated carbonic anhydrases and their inhibition. Curr Pharm Des. 14(7):685-98.
Gibadulinova A, Oveckova I, Parkkila S, Pastorekova S, Pastorek J. (2008) Key promoter elements involved in transcriptional activation of the cancer-related
gene coding for S100P calcium-binding protein. Oncology Reports, 20, 391-6.
Hilvo M, Baranauskiene L, Salzano AM, Scaloni A, Matulis D, Innocenti A, Scozzafava A, Monti SM, Di Fiore A, De Simone G, Lindfors M, Jänis J, Valjakka
J, Pastoreková S, Pastorek J, Kulomaa MS, Nordlund HR, Supuran CT, Parkkila S. (2008) Biochemical characterization of CA IX, one of the most
active carbonic anhydrase isozymes. J Biol Chem, 283(41), 27799-809.
Takacova M, Holotnakova T, Vondracek J, Machala M, Pencikova K, Gradin K, Poellinger L, Pastorek J, Pastorekova S, Kopacek J. Role of aryl
hydrocarbon receptor in modulation of the expression of the hypoxia marker carbonic anhydrase IX. Biochem J. 2009 Jan 20. [Epub ahead of print]
Pastorekova S, Pastorek J. Cancer-related carbonic anhydrase isozymes and their inhibition. In: Carbonic Anhydrase, its Inhibitors and Activators. Supuran
CT, Scozzafava A, Conway J, editors. CRC Press Boca Raton-London-New York-Washington D.C. 2004, p 253-279
WOS - CA IX papers and citations
Aplikovaný výskum – vstup komerčných firiem
od roku 1992 TRITON (USA)
CIBA CORNING corp. (USA)
CHIRON corp. (USA)
BAYER HEALTHCARE (USA)
BAYER SCHERING PHARMA (Germany)
SIEMENS (USA)
KONTRAKTY S FIRMAMI - CIBA, CHIRON, BAYER, WILEX
PATENTY
-
US patent s celosvetovou pôsobnosťou zameraný
na diagnostiku a terapiu
CENA VÝVOJA LIEČIVA a DIAGNOSTIKY
Firmy – vlastný vývoj
patenty
Akademické granty
WILEX
124I-cG250
PET/CT (coronal slice of the fused
abdominal image) in a patient with multiple
left renal masses. Uptake of antibody clearly
identifies the ccRCC (solid arrow); the mixed
cystic mass (dotted arrow) was not clear cell
phenotype.
FDG-PET in a patient with metastatic ccRCC.
The primary (solid arrow) is not clearly seen,
though the osseous (dashed arrow) and
nodal (dotted arrow) metastases are.
Signaling pathways as targets
for novel cancer therapies.
PI3K, phosphatidyl inositol-3kinase;
PTEN, phosphatase and
tensin homolog;
TSC, tuberous sclerosis
complex;
mTOR, mammalian target of
rapamycin;
HIF, hypoxiainducible
factor;
VHL, von Hippel–Lindau;
Cul, Cullin, ELO, elongins;
MAPK, mitogen-activated
protein kinase;
Hsp90, heat-shock protein 90;
VEGF, vascular endothelial
growth factor;
CXCR4, CXC chemokine
receptor-4;
EGFR, epidermal growth
factor receptor;
GLUT, glucose transporter.
AKÝ VÝSKUM ROBÍME?
ZÁKLADNÝ vs. APLIKOVANÝ (TRANSLAČNÝ)
From Wikipedia, the free encyclopedia
Applied research: is research accessing and using some part of the
research communities' (the academy's) accumulated theories, knowledge,
methods, and techniques, for a specific, often state, commercial, or client
driven purpose. Applied research is often opposed to pure research in
debates about research ideals, programs, and projects.
Basic research: (also called fundamental or pure research) has as its
primary objective the advancement of knowledge and the theoretical
understanding of the relations among variables (see statistics). It is
exploratory and often driven by the researcher’s curiosity, interest, and
intuition. Therefore, it is sometimes conducted without any practical end
in mind, although it may have confounding variables (unexpected
results) pointing to practical applications. The terms “basic” or
“fundamental” indicate that, through theory generation, basic research
provides the foundation for further, sometimes applied research. As
there is no guarantee of short-term practical gain, researchers may find
it difficult to obtain funding for basic research.
AKO DLHO TRVÁ TAKÝTO VÝSKUM/VÝVOJ
KTO HO PLATÍ
KOMU PATRIA VÝSLEDKY
NA VŠETKO TREBA PRINAJMENŠOM DVOCH
ĎAKUJEM ZA POZORNOSŤ
A ŽELÁM VÁM,
ABY NIKTO NEMUSEL POUŽIŤ NAŠE VÝSLEDKY