Journal reading

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Journal reading

報告者： PGY2 曾智皇報告日期 : 103.06.10 指導老師 : 林立民醫師陳玉昆醫師

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1. Introduction

2. CA-IX expression and prognosis in OSCC 3. CA-IX expression in other HNSCCs

4. Relationship between CA-IX and

chemoresistance or radioresistance

5. The relationship between CA-IX and other molecules

6. Role of CA-IX as therapeutic target against cancer

7. Conclusions

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1. Introduction

7. Conclusions

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Introduction

• Hypoxia and tumor cell proliferation

determine response to surgery, CT, and RT

• Tumor hypoxia 

a. The promotion of genetic instability b. The invasive capacity of tumor cells c. Metastasis

d. Worsening of the clinical evolution

 loss of the apoptotic capacity of cells

(5)

• Among the hypoxia markers, we will focus on the exogenous hypoxia markers, mainly 2-

nitroimidazole, pimonidazole, and EF-5

• In addition to exogenous markers, there are endogenous hypoxia markers, controlled by hypoxia-related genes, formed by HIF-1

(6)

• HIF-1 

a. Related to the von Hippel-Lindau (vHL) tumor suppressor protein during oncogenesis

b. Controls several target genes that involve the energy metabolism, angiogenesis (VEGF) and transmembrane carbonic anhydrases (CAs)

(7)

(8)

• CAs are transmembrane Zn metalo-enzymes

catalyze reversible hydration of carbon dioxide in carbonic acid

involved in respiration and acid-base equilibrium

• CA-IX and CA-XII are the two tumor-related carbonic anhydrases

(9)

• CA-IX is largely expressed in tumor cell lines and shows moderate-low expression in a

healthy gastrointestinal tract

• CA-IX expression has been thoroughly described in different tumors

 however, this has not been so for head and neck carcinomas

(10)

1. Introduction

7. Conclusions

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1. Introduction

7. Conclusions

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• Hoogsteen et al 2005 

a. Analyzed CA-IX expression and IdUrd in a series of 60 HNSCC cases, including 3 OSCC cases

b. Joint expression of CA-IX and IdUrd

responsible for repopulation and disease progression

(14)

• Le et al 2007

a. Elevated correlation of hypoxia levels and CA- IX

b. CA-IX  not related to any of the prognostic variables

(15)

• Kaanders et al 2002

a. Studied distribution of pimonidazole and CA- IX

b. Patients with hypoxic tumors and low

vascular density showed worse locoregional control

Although no relation was found between CA-IX expression and treatment outcome

(16)

• HNSCCs 

a. Hypoxic tumors by definition

b. Frequently diagnosed in very advanced stages

c. In HPV-positive cases that normally have better prognosis

(17)

• Brockton et al 2011

Showed that a high stromal expression of CA- IX is related to a reduced average survival in p16-negative tumors

• Kong et al 2009

a. 44% (36 of 82) HNSCC cases under study presented a strong HPV pyrosequencing signal

b. No relation with tumor hypoxia and CA-IX expression

(18)

1. Introduction

7. Conclusions

(19)

• Koukourakis et al 2001

a. CA-IX expression (26.6%, 20 of 75) takes place mainly in tumors with low

vascularization (CD-31), necrosis areas

b. CA-IX expression related to a poor overall response

 These results were confirmed by Jonathan et al and Beasley et al

(20)

• Bhattacharya et al 2004

a. HNSCC xenograf

b. Lack of microvessels in well-differentiated

areas of the xenograf related to hypoxia and positive for CA-IX

 a limited use of chemotherapeutic drugs  resistance to Irinotecan therapy

c. Hypoxia promoted the creation of resistant cell subpopulations

(21)

• Chintala et al 2010

a. The effect of Se-methylselenocysteine in HNSCC cell lines and xenografs

b. In cells and hypoxic areas, the combination of both drugs

 1. reduced HIF-1a levels 2. lowered CA-IX levels

(22)

• Zheng et al 2010

a. Transformed an OSCC  into PYM resistant and crossresistant to paclitaxel, Adriamycin, mitomycin

b. Application of CA inhibitor, acetazolamide, and CA-IX silencing with oligonucleotides  PH ↑ in resistant cells

 increase of chemosensitivity to PYM

(23)

Eriksen et al 2007

a. CA-IX as prognostic marker in a series of 320 HNSCCs undergoing radiotherapy +

nimorazole

b. CA-IX is not related to any clinical and pathological, prognostic parameters

(24)

• Patients treated with ARCON vs. conventional surgery ± radiotherapy

 the hypoxia and vascula density levels have no influence on treatment response

• Jonathan et al 2006

Relationship between CA-IX and the lack of locoregional control

disappears when tumors are treated with ARCON

*accelerated radiotherapy combined with carbogen and nicotinamide

(25)

Koukourakis et al 2006

Joint expression of HIF-2a and CA-IX is responsible for poor CHART results

* Continuous hyperfractionated accelerated radiotherapy

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1. Introduction

7. Conclusions

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HIF

• HIF-1a transcription is the regulating factor in cellular response to hypoxia

• In the case of OSCC, HIF-1a prevents apoptosis of tumor cells

• Zhu et al 2010 OSCC cell lines are cultivated with 1% O2 expression of mRNA CA-IX is

regulated by HIF-1a

(28)

• Chintala et al 2010

 Application of Se-methylselenocysteine combined with irinotecan

 HIF-1a ↓ CA-IX ↓

• Koukourakis et al

a. CA-IX expression  worse survival rates b. HIF-2a  worse locoregional control

c. Joint expression provoked an additive effect

 confirm relation between both markers

(29)

• Eckert et al 2010 HIF-1a & CA-IX

a. low expression of both proteins survived an average of 54.8 months

b. high HIF-1a and low CA-IX expression survived an average of 37.6 months;

tumor-related death risk was 4.97-fold

(30)

• Roh et al 2009

 Statistically significant relationship between CA-IX and HIF-1a and HIF-2a

• Winter et al

 Not confirm a positive correlation with HIF-1a nor HIF-2a

(31)

Ki-67

• Kim et al 2007 CA-IX & Ki-67

a. Correlation between both b. Established three patterns:

(1) high risk (elevated CA-IX ⁄ Ki-67)

(2) low risk (low CA-IX ⁄ Ki- 67)

(3) moderate risk (one of the two is elevated)

 The high-risk group was an independent prognostic factor for average survival and disease-free survival

(32)

• Kondo et al 2011

no relationship between Ki-67 expression and CA-IX

(33)

GLUT-1

• Schutter et al 2005 CA-IX & GLUT-1 (glucose transporter-1)

a. CA-IX expression was not related to 1. any of the clinical, pathological, or prognostic parameters

2. the expression of GLUT-1

• Kondo et al 2011 & Jonathan et al 2006

 not found relation between GLUT-1 expression and CA-IX

(34)

Erythropoietin (Epo) and erythropoietin receptor (EpoR)

• Winter et al 2005

a. Tried to link tumor hypoxia and its main marker, CA-IX, with Hb, Epo, and EpoR

b. Positive correlation in CA-IX and Epo, but not EpoR

c. No correlation between Hb and Epo or EpoR

(35)

Other molecules

• Le et al 2007

significant relationship between intense CA-IX expression and hypoxia-related proteins:

BNIP3L, LOX, CTGF, Ephrin A1, GAL-1

(36)

• Le et al 2007

Positive correlation between the expression of CA-IX and Galectin-1 and their relation with

treatment outcome

• Silva et al 2010

Co-expression of CA-IX and MVP (major vault protein) confers tumors a significantly lesser chance of locoregional control

(37)

• Gee et al 2010

a. Abnormally high levels of microRNA hsa-miR- 210 in 46 HNSCC patients

b. Statistically significant relationship between hypoxia markers such as HIF-1a, the TWIST1 gene, and CA-IX

c. Locoregional recurrence with a smaller average survival

(38)

• Schutter et al 2006

a. Analyzing the relation between micro- satellite alterations and HNSCC tumor hypoxia

b. LOH (loss of heterozygosity) is very frequent in regions close to p53, specifically in

D17S799, in patients that simultaneously showed high CA-IX expression

supporting the theory of the relationship between p53 and hypoxia

(39)

1. Introduction

7. Conclusions

(40)

• CA-IX and XII are predominantly found in tumor cells

• Hydrating carbon dioxide to protons and

bicarbonate, these CAs contribute significantly to the extracellular acidification of solid

tumors

• CA-IX is overexpressed in many tumors in response to the HIF pathway

(41)

• Design campaigns of inhibitors against this novel, recently validated antitumor target

• Obtaining compounds that specifically target the tumor-associated isoforms CA-IX and XII

• Coumarin and thiocoumarins are the most important such new CAIs

(42)

Lou Y et al 2011

Pacchiano F et al 2011 

• The most interesting CAIs reported to date are the ureido-sulfonamide and the glycosyl

coumarin (low nanomolar CA-IX selective inhibitors)



which strongly inhibit the growth of both primary tumors and metastases in several animal models of breast cancer

(43)

• Similar animal model of breast cancer cell lines which does not express CA-IX has been used as negative control

 no effects on the growth of the tumors have been evidenced afer treatment with sulfonamide ⁄ coumarin CAIs

(44)

 demonstrated the potential of CA-IX inhibition for designing antitumor ⁄

antimetastatic agents possessing a novel mechanism of action

(45)

Zatovicova M et al 2010 

• M75 is a highly specific anti-CA-IX mAb targeting the PG domain of CA-IX

used in immunohistochemical and western blot studies for evidencing CA-IX

(46)

Siebels M et al 2011 

• WX-G250 (Girentuximab) is another anti-CA-IX chimeric monoclonal antibody in phase III

clinical trials as adjuvant therapy for the treatment of non-metastasized RCC

(47)

• Tumor-associated CAs are indeed almost ideal targets for designing novel and innovative

anticancer drugs

• Interfere with tumor acidification by a

mechanism of action not yet exploited by the classical cytostatic drugs

(48)

1. Introduction

7. Conclusions

(49)

• Hypoxia in solid tumors is a decisive factor for the outcome of HNSCCs

• Hypoxia in solid tumors with chemoresistance and the failure of radiotherapy, both

conventional and concomitant, in which CA-IX seems to play an important role

(50)

• Several mAbs (girentuximab, and its 124I- radiolabelled variant) targeting CA-IX

 in advanced clinical trials both for a. treatment

b. imaging of hypoxic tumors overexpressing this enzyme

(51)

• Two small molecule CA-IX inhibitors  1. sulfonamide

2. glycosyl coumarin

 both for imaging and treatment of solid tumors and metastases in which CA-IX is present

(52)

• Further studies of these tumors are needed to confirm

 a. the use of CA-IX as a prognostic marker b. evaluate its possible inhibition with

minimal adverse effects

c. reducing the risk of metastasis

d. favoring the action of chemotherapeutic drugs and radiotherapy

(53)

Journal reading

Journal reading

Introduction

HIF

Ki-67

GLUT-1

Erythropoietin (Epo) and erythropoietin receptor (EpoR)

Other molecules

Thank you for your

attention