Conversion of a tumor-binding peptide identified by phage display to a functional chimeric T cell antigen receptor

Pameijer, Colette R.; Navanjo, A.; Meechoovet, Bessie; Wagner, Jamie R.; Aguilar, Brenda; Wright, Chris J.; Chang, Wen Chung; Brown, Christine E.; Jensen, Michael C.

doi:10.1038/sj.cgt.7700993

Cited by 53 publications

(31 citation statements)

References 13 publications

(17 reference statements)

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“…The mimicry phenomenon for peptides isolated from phage-display libraries can be investigated on many levels including phage-expressed and free peptides, or peptides fused to proteins (31,32). Finally, characterization of functional (agonist or antagonist activity) or immunological mimicry of peptides can be performed (33,34). We aimed to probe the molecular basis of mAb 14G2a binding by peptides isolated from the LX-8 library.…”

Section: Discussionmentioning

confidence: 99%

Analysis and optimization of interactions between peptides mimicking the GD2 ganglioside and the monoclonal antibody 14G2a

Horwacik

Kurciński²,

Bzowska³

et al. 2011

Int J Mol Med

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Abstract. Overexpression of the Gd2 ganglioside (Gd2) is a hallmark of neuroblastoma. The antigen is used in neuroblastoma diagnosis and to target newly developed therapies to cancer cells. Peptide mimetics are novel approaches in the design of antigens for vaccine development. We previously reported the isolation of five GD2-mimicking peptides from the LX-8 phage display library with the monoclonal antibody (mAb) 14G2a. The goal of our current study was to analyze and optimize the binding of the peptide mimetics to the mAb 14G2a. Therefore, we performed further experiments and supported them with molecular modeling to investigate structure-activity relationships that are the basis for the observed mimicry of Gd2 by our peptides. Here, we show that the peptides have overlapping binding sites on the mAb, 14G2a and restricted specificity, as they did not crossreact with other ganglioside-specific antibodies tested. In addition we demonstrate that the phage environment was involved in the process of selection of our peptides. The AAEGd sequence taken from the viral major coat protein, p8, and added to the c-termini of the peptides #65, #85 and #94 significantly improved their binding to the mAb, 14G2a. By application of analogs with amino acid substitutions and sequence truncations, we elucidated the structure-activity relationships necessary for the interactions between the 14G2a mAb and the peptide #94 (RCNPNMEPPRCF). We identified amino acids indispensable for the observed Gd2-mimicry by #94 and confirmed a pivotal role of the disulphide bridge between the cysteine residues of #94 for binding to the mAb 14G2a. More importantly, we report five new peptides demonstrating a significant improvement of mAb 14G2a binding. The experimental data were supported and expanded with molecular modeling tools. Taken together, the experimental results and the in silico data allowed us to probe in detail the mechanism of the molecular mimicry of Gd2 by the peptides. Additionally, we significantly optimized binding of the leading peptide sequence #94 to the mAb 14G2a. We can conclude that our findings add to the knowledge on factors governing selections of peptide mimetics from phage-display libraries.

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Section: Discussionmentioning

confidence: 99%

Analysis and optimization of interactions between peptides mimicking the GD2 ganglioside and the monoclonal antibody 14G2a

Horwacik

Kurciński²,

Bzowska³

et al. 2011

Int J Mol Med

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“…Similarly, significant amounts of IFN-γ were released in the presence of αvβ 6-positive tumor cells and this release could be blocked by the addition of soluble tumor-binding peptide, but not an irrelevant peptide [51].…”

Section: Peptides As Car-binding Domainsmentioning

confidence: 93%

“…For example, Pameijer and colleagues took advantage of a tumor-binding peptide specific for integrin alpha vβ 6 that was discovered by phage display [51]. Alpha vβ 6 integrin is overexpressed on squamous cell carcinomas of the head and neck, cervix, skin and esophagus, but has a limited and low level expression on normal tissues [52].…”

Section: Peptides As Car-binding Domainsmentioning

confidence: 99%

Foreign or Domestic CARs: Receptor Ligands as Antigen-Binding Domains

Shaffer

Zhou

2014

Medical Sciences

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Chimeric antigen receptors (CARs) are increasingly being used in clinical trials to treat a variety of malignant conditions and recent results with CD19-specific CARs showing complete tumor regressions has sparked the interest of researchers and the public alike. Traditional CARs have been generated using single-chain variable fragments (scFv), often derived from murine monoclonal antibodies, for antigen specificity. As the clinical experience with CAR T cells grows, so does the potential for unwanted immune responses against the foreign transgene. Strategies that may reduce the immunogenicity of CAR T cells are humanization of the scFv and the use of naturally occurring receptor ligands as antigen-binding domains. Herein, we review the experience with alternatively designed CARs that contain receptor ligands rather than scFv. While most of the experiences have been in the pre-clinical setting, clinical data is also emerging.

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“…Reports demonstrate the identification of mHAs or tumor antigens using serologic expression methodologies like SEREX 22,23 or chimeric antigen receptors of tumor-specific T lymphocytes coupled with phage display library 24 after transplantations. We hypothesized that novel mHAs could be serologically identified as targets of antibody responses that develop after allogeneic transplantations and are absent before transplantation.…”

Section: Discussionmentioning

confidence: 99%

Antibodies specifically target AML antigen NuSAP1 after allogeneic bone marrow transplantation

Wadia

Coram

Armstrong

et al. 2010

Blood

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Identifying the targets of immune response after allogeneic hematopoietic cell transplantation (HCT) promises to provide relevant immune therapy candidate proteins. We used protein microarrays to serologically identify nucleolar and spindleassociated protein 1 (NuSAP1) and chromatin assembly factor 1, subunit B (p60; CHAF1b) as targets of new antibody responses that developed after allogeneic HCT. Western blots and enzyme-linked immunosorbent assays (ELISA) validated their post-HCT recognition and enabled ELISA testing of 120 other patients with various malignancies who underwent allo-HCT. CHAF1b-specific antibodies were predominantly detected in patients with acute myeloid leukemia (AML), whereas NuSAP1-specific antibodies were exclusively detected in patients with AML 1 year after transplantation (P < .001).

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Conversion of a tumor-binding peptide identified by phage display to a functional chimeric T cell antigen receptor

Cited by 53 publications

References 13 publications

Analysis and optimization of interactions between peptides mimicking the GD2 ganglioside and the monoclonal antibody 14G2a

Analysis and optimization of interactions between peptides mimicking the GD2 ganglioside and the monoclonal antibody 14G2a

Foreign or Domestic CARs: Receptor Ligands as Antigen-Binding Domains

Antibodies specifically target AML antigen NuSAP1 after allogeneic bone marrow transplantation

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