CXCR4 receptor is correlated to the development of metastases in leiomyosarcoma, pleomorphic sarcoma and

liposarcoma

S.C.D.U. Medical Oncology S.C.D.U. Medical Oncology I.R.C.C. CandioloI.R.C.C. Candiolo

Ordine MaurizianoOrdine Mauriziano

S Aliberti, G Grignani, G Cavalloni, A Pisacane, P Allione,I Sarotto, Y Pignochino, M Motta, B Torchio, M Risio, M Aglietta.

“When a plant goes to seed, its seeds are carried in all directions; but they can only live and grow if they fall on congenial soil.”

S Paget The Lancet 1889

-3149 adult patients with soft tissue sarcoma were admitted and treated at Memorial Sloan-Kettering Cancer Center.

-719 patients developed lung metastases (about 24%)

Tumor progression steps

Cancer cell

YY

chemokine receptor

chemokine

endothelium

Non-target tissue

Cancer cell

Y Yendothelium

differentchemokine

Target tissue

Pancreatic cancer Kosshiba et al., Clin. Cancer Res. 2000

Neuroblastoma Geminder et al., J. Immunol. 2001

Breast cancer Muller et al., Nature 2001

Rhabdomyosarcoma Libura et al., Blood 2002

Prostate cancer Taichman et al., Cancer Res. 2002

Colon cancer Zeelemberg et al., Cancer Res. 2003

Osteosarcoma Perissinotto et al., Clin Cancer Res. 2005

Laverdiere et al., Clin Cancer Res. 2005

Non-small cell lung cancer cells Su et al., Clin Cancer Res. 2005

Esophageal cancer Kaifi et al., J Natl Cancer Inst. 2005

Carcinomatosis of Gastric Cancer Yasumoto et al., Cancer Res. 2006

Glioma Ehtesham et al., Oncogene 2006

Melanoma Bartolome et al., Cancer Res. 2006

Involvement of CXCR4/SDF-1 system in cancer progression

Purpose

• To analyze expression patterns of CXCR4, EGFR, angioinvasiveness (by standard immunohistochemical technique) along with other primary tumor features (T and grading).

• To assess if there is a correlation among the characteristics of primary tumor and the development of metastases.

Materials and methods

We retrospectively evaluated expression patterns ofCXCR4, EGFR and angioinvasiveness features onprimary tumors in 67 pts.

-Mean age = 57 yrs (24 - 85)-Male/female = 38/29-Median follow-up = 57 mos (2 – 160)-Limbs/retroperitoneal = 47/20-Histotype: leiomyosarcoma, pleomorphic sarcoma

non-myxoid liposarcoma.-Grading: according to Coindre et al.

Materials and methods

Site N°

Lung 13 40%

Lung + other

11 18%

Liver 6 18%

Other 3 9%

CXC4 immunohistochemistry

CXCR4: Pharmagin 12G-5 monoclonal antibody

Cytoplasmic positivity Nuclear positivity

EGFR immunohistochemistry

EGFR: pharmaDx monoclonal antibody DakoCytomation

Positive = 3+ & 80% of cells Negative = 0/1+ & < 1% of cells

Angioinvasiveness features

Results

Negative 34/67

50.7%

Positive 33/67

49.3%

Negative 29/67

43.2%

Low 19/67

28.4%

High 19/67

28.4%

Negative 13/67

19.4%

Low 12/67

17.9%

High 42/67

62.7%

CXCR4 EGFRAngio-

Invasiveness

Variable

CXCR4

EGFR

Angiogenesis

Grading

T

Tumor burden

0.07

0.05

0.018

0.32

0.017

Univariate

0.067

0.03

0.06

0.02

Multivariate

Correlation with development of metastases

Response according to metastasisCumulativ e Proportion Surv iv ing (Kaplan-Meier)

Complete Cens ored

0 50 100 150

Time

0,2

0,3

0,4

0,5

0,6

0,7

0,8

0,9

1,0

Cum

ulative Proportion S

urviving

p = .00645

CXCR4 ne g

CXCR4 pos

Cumulativ e Proportion Surv iv ing (Kaplan-Meier)

Complete Cens ored

0 20 40 60 80 100 120

Time

0,0

0,1

0,2

0,3

0,4

0,5

0,6

0,7

0,8

0,9

1,0

Cum

ulative Proportion S

urviving

CXCR4 - stratified by grading

CXCR4 pos p = 0.018

CXCR4 neg

Cumulativ e Proportion Surv iv ing (Kaplan-Meier)

Complete Cens ored

0 20 40 60 80 100 120

Time

0,4

0,5

0,6

0,7

0,8

0,9

1,0

Cum

ulative Proportion S

urviving

CXCR4 neg

CXCR4 pos

p = 0.05

Grade = 1 or 2 Grade = 3

Where do we go from here?

CXCR4 expression on leiomyosarcoma cell lines

Cell line % CXCR4+ cells

SK-LMS-1

Leiomyosarcoma

20-70%

SK-UT-1

Leiomyosarcoma

20-70%

MES-SA Uterine leiomyosarcoma

10-12%

91266 Uterine leiomyosarcoma

Negative

0

5

10

15

20

25

30

35

40

45

SK-LMS-1-25 SK-LMS-1-50 SK-UT-1-25 SK-UT-1-50

NT SDF-1 1 h 100 ng ml

CXCR4 down-regulation by SDF-1 on CXCR4+ leiomyosarcoma cell lines

Regulation of CXCR4 expression by in vitro gene transduction in leiomyosarcoma cell line

AMD3465 selectively reduces CXCR4 transcript expression in lungs.

• In this retrospective series we show how CXCR4 is associated with the development of lung metastases.

• We are currently verifying these results on larger series.

• We are developing an in vitro model in which we can modulate CXCR4 expression.

• The metastatic behavior of these histotypes needs further study but we believe CXCR4 may play a role in the recurrence of soft tissue sarcoma.

Conclusions

Acknowledgments

Medical Oncology:

Surgical Pathology:

Surgical Pathology:

Sandra AlibertiPaolo AllioneMassimo Aglietta

Alberto PisacaneIvana SarottoMauro Risio

Manuela MottaBruno Torchio

Molecular Oncology: Giuliana CavalloniYmera Pignochino