Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery

J. Posthuma de Boer; S.R. Piersma; T.V. Pham; P.W. van Egmond; J.C. Knol; A.M. Cleton-Jansen; M.A. van Geer; V.W. van Beusechem; G.J.L. Kaspers; B.J. van Royen; C.R. Jimenez; M.N. Helder; J. PosthumaDeBoer

doi:https://doi.org/10.1038/bjc.2013.578

Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery

J. Posthuma de Boer, S.R. Piersma, T.V. Pham, P.W. van Egmond, J.C. Knol, A.M. Cleton-Jansen, M.A. van Geer, V.W. van Beusechem, G.J.L. Kaspers, B.J. van Royen, C.R. Jimenez, M.N. Helder, J. PosthumaDeBoer

Research output: Contribution to journal › Article › Academic › peer-review

42 Citations (Scopus)

Abstract

Background:Osteosarcoma (OS) is the most common bone tumour in children and adolescents. Despite aggressive therapy regimens, treatment outcomes are unsatisfactory. Targeted delivery of drugs can provide higher effective doses at the site of the tumour, ultimately improving the efficacy of existing therapy. Identification of suitable receptors for drug targeting is an essential step in the design of targeted therapy for OS.Methods:We conducted a comparative analysis of the surface proteome of human OS cells and osteoblasts using cell surface biotinylation combined with nano-liquid chromatography - tandem mass spectrometry-based proteomics to identify surface proteins specifically upregulated on OS cells. This approach generated an extensive data set from which we selected a candidate to study for its suitability as receptor for targeted treatment delivery to OS. First, surface expression of the ephrin type-A receptor 2 (EPHA2) receptor was confirmed using FACS analysis. Ephrin type-A receptor 2 expression in human tumour tissue was tested using immunohistochemistry. Receptor targeting and internalisation studies were conducted to assess intracellular uptake of targeted modalities via EPHA2. Finally, tissue micro arrays containing cores of human OS tissue were stained using immunohistochemistry and EPHA2 staining was correlated to clinical outcome measures.Results:Using mass spectrometry, a total of 2841 proteins were identified of which 156 were surface proteins significantly upregulated on OS cells compared with human primary osteoblasts. Ephrin type-A receptor 2 was highly upregulated and the most abundant surface protein on OS cells. In addition, EPHA2 was expressed in a vast majority of human OS samples. Ephrin type-A receptor 2 effectively mediates internalisation of targeted adenoviral vectors into OS cells. Patients with EPHA2-positive tumours showed a trend toward inferior overall survival.Conclusion:The results presented here suggest that the EPHA2 receptor can be considered an attractive candidate receptor for targeted delivery of therapeutics to OS

Original language	English
Pages (from-to)	2142-2154
Journal	British journal of cancer
Volume	109
Issue number	8
DOIs	https://doi.org/10.1038/bjc.2013.578
Publication status	Published - 2013

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https://doi.org/10.1038/bjc.2013.578

Cite this

Posthuma de Boer, J., Piersma, S. R., Pham, T. V., van Egmond, P. W., Knol, J. C., Cleton-Jansen, A. M., van Geer, M. A., van Beusechem, V. W., Kaspers, G. J. L., van Royen, B. J., Jimenez, C. R., Helder, M. N., & PosthumaDeBoer, J. (2013). Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery. British journal of cancer, 109(8), 2142-2154. https://doi.org/10.1038/bjc.2013.578

@article{ae7a8afb6b784c1e9e1ed2a89b1b8013,

title = "Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery",

abstract = "Background:Osteosarcoma (OS) is the most common bone tumour in children and adolescents. Despite aggressive therapy regimens, treatment outcomes are unsatisfactory. Targeted delivery of drugs can provide higher effective doses at the site of the tumour, ultimately improving the efficacy of existing therapy. Identification of suitable receptors for drug targeting is an essential step in the design of targeted therapy for OS.Methods:We conducted a comparative analysis of the surface proteome of human OS cells and osteoblasts using cell surface biotinylation combined with nano-liquid chromatography - tandem mass spectrometry-based proteomics to identify surface proteins specifically upregulated on OS cells. This approach generated an extensive data set from which we selected a candidate to study for its suitability as receptor for targeted treatment delivery to OS. First, surface expression of the ephrin type-A receptor 2 (EPHA2) receptor was confirmed using FACS analysis. Ephrin type-A receptor 2 expression in human tumour tissue was tested using immunohistochemistry. Receptor targeting and internalisation studies were conducted to assess intracellular uptake of targeted modalities via EPHA2. Finally, tissue micro arrays containing cores of human OS tissue were stained using immunohistochemistry and EPHA2 staining was correlated to clinical outcome measures.Results:Using mass spectrometry, a total of 2841 proteins were identified of which 156 were surface proteins significantly upregulated on OS cells compared with human primary osteoblasts. Ephrin type-A receptor 2 was highly upregulated and the most abundant surface protein on OS cells. In addition, EPHA2 was expressed in a vast majority of human OS samples. Ephrin type-A receptor 2 effectively mediates internalisation of targeted adenoviral vectors into OS cells. Patients with EPHA2-positive tumours showed a trend toward inferior overall survival.Conclusion:The results presented here suggest that the EPHA2 receptor can be considered an attractive candidate receptor for targeted delivery of therapeutics to OS",

author = "{Posthuma de Boer}, J. and S.R. Piersma and T.V. Pham and {van Egmond}, P.W. and J.C. Knol and A.M. Cleton-Jansen and {van Geer}, M.A. and {van Beusechem}, V.W. and G.J.L. Kaspers and {van Royen}, B.J. and C.R. Jimenez and M.N. Helder and J. PosthumaDeBoer",

year = "2013",

doi = "https://doi.org/10.1038/bjc.2013.578",

language = "English",

volume = "109",

pages = "2142--2154",

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issn = "0007-0920",

publisher = "Nature Publishing Group",

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Posthuma de Boer, J, Piersma, SR , Pham, TV, van Egmond, PW, Knol, JC, Cleton-Jansen, AM, van Geer, MA, van Beusechem, VW , Kaspers, GJL , van Royen, BJ , Jimenez, CR , Helder, MN & PosthumaDeBoer, J 2013, 'Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery', British journal of cancer, vol. 109, no. 8, pp. 2142-2154. https://doi.org/10.1038/bjc.2013.578

TY - JOUR

T1 - Surface proteomic analysis of osteosarcoma identifies EPHA2 as receptor for targeted drug delivery

AU - Posthuma de Boer, J.

AU - Piersma, S.R.

AU - Pham, T.V.

AU - van Egmond, P.W.

AU - Knol, J.C.

AU - Cleton-Jansen, A.M.

AU - van Geer, M.A.

AU - van Beusechem, V.W.

AU - Kaspers, G.J.L.

AU - van Royen, B.J.

AU - Jimenez, C.R.

AU - Helder, M.N.

AU - PosthumaDeBoer, J.

PY - 2013

Y1 - 2013

N2 - Background:Osteosarcoma (OS) is the most common bone tumour in children and adolescents. Despite aggressive therapy regimens, treatment outcomes are unsatisfactory. Targeted delivery of drugs can provide higher effective doses at the site of the tumour, ultimately improving the efficacy of existing therapy. Identification of suitable receptors for drug targeting is an essential step in the design of targeted therapy for OS.Methods:We conducted a comparative analysis of the surface proteome of human OS cells and osteoblasts using cell surface biotinylation combined with nano-liquid chromatography - tandem mass spectrometry-based proteomics to identify surface proteins specifically upregulated on OS cells. This approach generated an extensive data set from which we selected a candidate to study for its suitability as receptor for targeted treatment delivery to OS. First, surface expression of the ephrin type-A receptor 2 (EPHA2) receptor was confirmed using FACS analysis. Ephrin type-A receptor 2 expression in human tumour tissue was tested using immunohistochemistry. Receptor targeting and internalisation studies were conducted to assess intracellular uptake of targeted modalities via EPHA2. Finally, tissue micro arrays containing cores of human OS tissue were stained using immunohistochemistry and EPHA2 staining was correlated to clinical outcome measures.Results:Using mass spectrometry, a total of 2841 proteins were identified of which 156 were surface proteins significantly upregulated on OS cells compared with human primary osteoblasts. Ephrin type-A receptor 2 was highly upregulated and the most abundant surface protein on OS cells. In addition, EPHA2 was expressed in a vast majority of human OS samples. Ephrin type-A receptor 2 effectively mediates internalisation of targeted adenoviral vectors into OS cells. Patients with EPHA2-positive tumours showed a trend toward inferior overall survival.Conclusion:The results presented here suggest that the EPHA2 receptor can be considered an attractive candidate receptor for targeted delivery of therapeutics to OS

AB - Background:Osteosarcoma (OS) is the most common bone tumour in children and adolescents. Despite aggressive therapy regimens, treatment outcomes are unsatisfactory. Targeted delivery of drugs can provide higher effective doses at the site of the tumour, ultimately improving the efficacy of existing therapy. Identification of suitable receptors for drug targeting is an essential step in the design of targeted therapy for OS.Methods:We conducted a comparative analysis of the surface proteome of human OS cells and osteoblasts using cell surface biotinylation combined with nano-liquid chromatography - tandem mass spectrometry-based proteomics to identify surface proteins specifically upregulated on OS cells. This approach generated an extensive data set from which we selected a candidate to study for its suitability as receptor for targeted treatment delivery to OS. First, surface expression of the ephrin type-A receptor 2 (EPHA2) receptor was confirmed using FACS analysis. Ephrin type-A receptor 2 expression in human tumour tissue was tested using immunohistochemistry. Receptor targeting and internalisation studies were conducted to assess intracellular uptake of targeted modalities via EPHA2. Finally, tissue micro arrays containing cores of human OS tissue were stained using immunohistochemistry and EPHA2 staining was correlated to clinical outcome measures.Results:Using mass spectrometry, a total of 2841 proteins were identified of which 156 were surface proteins significantly upregulated on OS cells compared with human primary osteoblasts. Ephrin type-A receptor 2 was highly upregulated and the most abundant surface protein on OS cells. In addition, EPHA2 was expressed in a vast majority of human OS samples. Ephrin type-A receptor 2 effectively mediates internalisation of targeted adenoviral vectors into OS cells. Patients with EPHA2-positive tumours showed a trend toward inferior overall survival.Conclusion:The results presented here suggest that the EPHA2 receptor can be considered an attractive candidate receptor for targeted delivery of therapeutics to OS

U2 - https://doi.org/10.1038/bjc.2013.578

DO - https://doi.org/10.1038/bjc.2013.578

M3 - Article

C2 - 24064975

SN - 0007-0920

VL - 109

SP - 2142

EP - 2154

JO - British journal of cancer

JF - British journal of cancer

IS - 8

ER -