Phage display selection of peptides that affect prostate carcinoma cells attachment and invasion

Romanov, Victor; Durand, D; Petrenko, Valery A.

doi:10.1002/pros.1068

Cited by 73 publications

(37 citation statements)

References 33 publications

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“…Biopanning was performed as described previously (11). The ratio of output:input (number of cell-associated phage divided by the numbers of total phage applied to the cells) was calculated for each round.…”

Section: Introductionmentioning

confidence: 99%

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Prostate Cancer Cell Adhesion to Bone Marrow Endothelium

Romanov¹,

Whyard²,

Adler³

et al. 2004

Cancer Research

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Bone metastasis is the most frequent complication of prostate cancer (PC). Elucidation of the biological basis of this specificity is required for the development of approaches for metastatic inhibition. We investigated the possibility that the preferential attachment of PC cells to bone marrow endothelium (as opposed to endothelium from other organs) affects this specificity. We selected, from peptide phage-displayed libraries, peptide ligands to surfaces of PC cells (C4-2B) attenuated (30 -40%) binding of C4-2B cells to bone marrow endothelial cells (BMECs). We then determined the molecules on the surface of C4-2B cells interacted with the selected peptides using column affinity chromatography and a cDNA expression phage-displayed library generated from C4-2B cells in T7 phage. We identified a phage from the cDNA library that specifically bound to one of the selected peptides-L11. This phage displayed the amino acid sequence homologous for the COOH-terminal portion of prostatespecific antigen (PSA). To examine the possible direct involvement of PSA in the interactions between PC and BMECs, we performed a cell-cell adhesion assay. Antibodies to PSA attenuated PC cells adhesion to BMECs. In addition, exogenous proteolytically active PSA modulated this adhesion. Finally, inactivation of mRNA coding PSA by a small interfering RNA (siRNA) diminished C4-2B cell adhesion to BMECs. These results indicate that PSA expressed as secreted and surface-associated molecules in C4-2B cells is involved in cell-cell interactions and/or digests components of bone marrow endothelium for preferential adhesion and penetration of PC cells. The suggested experimental approach is a promising strategy for identification of cell surface molecules involved in intercellular interactions.

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Section: Introductionmentioning

confidence: 99%

“…The final round of biopanning was defined after this parameter reached the value of 0.5-1.0%. The sequences of DNA coding inserts were determined for 48 of the selected phage from the output of the last round of biopanning as described previously (11). Translated peptide sequences were aligned for further analysis.…”

Section: Introductionmentioning

confidence: 99%

Prostate Cancer Cell Adhesion to Bone Marrow Endothelium

Romanov¹,

Whyard²,

Adler³

et al. 2004

Cancer Research

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“…30,35 Resulting peptide array is uniformly spaced at 2.7 nm apart 30 and is readily available for cell receptor interaction. 36 Thus by cloning an insert at this location, we constructed a nanofilament with a very dense display of signaling peptides (1.5 × 10 13 epitopes/cm 2 ) (calculations shown in Supporting Information). The peptides chosen to be displayed on the phage were RGD, a cell adhesion integrin binding motif, and IKVAV, a laminin motif known to promote neural cell adhesion and neurite extension.…”

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confidence: 99%

Genetically Engineered Nanofiber-Like Viruses For Tissue Regenerating Materials

2009

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Controlling structural organization and signaling motif display of biomimetic matrices at the nanometer scale is of great importance to the functional design of tissue regenerating materials. We have genetically engineered M13 bacteriophage (phage), naturally occurring nanofiberlike viruses, to display a high density of cell-signaling peptides on their major coat proteins. Structural orientation of these phage building blocks can be achieved due to their long-rod shape and monodispersity, which lead them to self-assemble into directionally organized liquid crystalline-like materials. We showed that the constructed viral nanofiber scaffolds were able to support neural progenitor cell proliferation and differentiation as well as direct orientation of their growth in three dimensions. Such functionalized and structurally aligned phage matrices offer promising opportunities for therapies that address challenging medical problems, such as nerve tissue regeneration after spinal cord injuries, or as in vitro model systems for studying complicated cell signaling environments.Designing biomimetic matrices with a precise control of structural organization and display of signaling motifs to regulate cell behavior is one of the most critical issues for the development of tissue regenerating materials. In nature, a nanofibrous protein network, in combination with neighboring cells, support and guide cellular behavior through a multitude of chemical and physical cues. [1][2][3][4] Similarly, manmade materials are being developed to recreate an environment that would provide for comparable controls over cell activities. [5][6][7][8][9][10][11] Recently developed top-down and bottomup material synthesis approaches 6,[8][9][10][11][12][13][14][15] are allowing for biomimetic materials to approximate a level of complexity observed in the natural extracellular environment. Nanofibers offer increased surface area for cell engagement, 10,13 which has led to improved cell attachment and material integration, as well as enhanced cell proliferation and differentiation. 6,12,13 Controlling the density of displayed signaling molecules at the nanometer scale has permitted quantitative investigations of cell behaviors. 9,11,12,[16][17][18] Observations of cell growth on substrates with aligned topographical features, such as nanofibers or ridges, have verified that cells can sense and respond to ordered nanoscale textures with polarization and coalignment. 6,13,19 Furthermore, self-assembling peptide-based materials have been utilized for convenient cell entrapment in three dimensions and offer attractive opportunities for minimally invasive therapy applications. 12,17 These various engineering approaches provide different combinations of advantageous material attributes, yet no single technique has been able to simultaneously satisfy the following multiple requirements of tissue regenerating scaffolds: to provide a cell conducive environment, to display signaling molecules in a controlled manner, to form self-organized nanofilamen...

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“…Phages can be selected from landscape phage display libraries with affinities for a wide range of simple targets such as dioxin, Cibacron blue, β-galactosidase, streptavidin, neutravidin and fibrinogen [11,13,14,27], as well as for more complex targets such as prostate cancer cells [15,16,28], malignant glial cells [17,25,30], or serum antibodies [31]. Landscape phages have been shown to serve as substitutes for antibodies and detection probes in enzyme-linked immuno assay (ELISA) and biosensors [13][14][15][16][17][18][19][20][21][22][23], as immunogens [24], gene-delivery vehicles [25] and affinity matrices [30,32].…”

Section: Landscape Phages As Substitute Antibodiesmentioning

confidence: 99%

“…Landscape phages have been shown to serve as substitutes for antibodies and detection probes in enzyme-linked immuno assay (ELISA) and biosensors [13][14][15][16][17][18][19][20][21][22][23], as immunogens [24], gene-delivery vehicles [25] and affinity matrices [30,32].…”

Section: Landscape Phages As Substitute Antibodiesmentioning

confidence: 99%

Landscape phage as a molecular recognition interface for detection devices

Petrenko

2008

Microelectronics Journal

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Filamentous phages are thread-shaped bacterial viruses. Their outer coat is a tube formed by thousands equal copies of the major coat protein pVIII. Libraries of random peptides fused to pVIII domains were used for selection of phages probes specific for a panel of test antigens and biological threat agents. Because the viral carrier in the phage borne bio-selective probes is infective, they can be cloned individually and propagated indefinitely without needs of their chemical synthesis or reconstructing. As a new bioselective material, landscape phages combine unique characteristics of affinity reagents and self assembling proteins. Biorecognition layers formed by the phage-derived probes bind biological agents with high affinity and specificity and generate detectable signals in analytical platforms. The performance of phage-derived materials as biorecognition interface was illustrated by detection of Bacillus anthracis spores and Salmonella typhimurium cells. With further refinement, the phage-derived analytical platforms for detecting and monitoring of numerous threat agents may be developed, since phage interface against any bacteria, virus or toxin may be readily selected from the landscape phage libraries. As an interface in the field-use detectors, they may be superior to antibodies, since they are inexpensive, highly specific and strong binders, resistant to high temperatures and environmental stresses.

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Phage display selection of peptides that affect prostate carcinoma cells attachment and invasion

Abstract: These results indicate that identified ligands may play a role in tumorigenicity and metastatic transformation of prostate cancer. To our knowledge, this is the first identification of a functional cancer-specific peptide ligand using the phage display approach.

Cited by 73 publications

References 33 publications

Prostate Cancer Cell Adhesion to Bone Marrow Endothelium

Prostate Cancer Cell Adhesion to Bone Marrow Endothelium

Genetically Engineered Nanofiber-Like Viruses For Tissue Regenerating Materials

Landscape phage as a molecular recognition interface for detection devices

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