High-affinity peptide against MT1-MMP for in vivo tumor imaging

Zhu, Lei; Wang, Huiling; Wang, Lin; Wang, Ye; Jiang, Kun; Li, Cheng; Ma, Qingjie; Gao, Shi; Wang, Liping; Li, Wei; Cai, Mingjun; Wang, Hongda; Niu, Gang; Lee, Seulki; Yang, Wei; Fang, Xuexun; Chen, Xiaoyuan

doi:10.1016/j.jconrel.2011.01.032

Cited by 58 publications

(58 citation statements)

References 41 publications

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“…Because existing methods are already capable of discerning strong from weak and non-binders, we have focused on discriminating tight binders within a 500-fold range of affinities (0.1 nM-60 nM). Accurate data in this regime may aid in the design of very strong binders that can be important therapeutic and diagnostic agents [17–19]. We conducted our study using a small library of about 1000 yeast displayed BH3 peptides that bind to Bcl - x L , a key regulator of apoptosis.…”

Section: Introductionmentioning

confidence: 99%

SORTCERY—A High–Throughput Method to Affinity Rank Peptide Ligands

Reich

Dutta

Keating

2015

Journal of Molecular Biology

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Uncovering the relationships between peptide and protein sequences and binding properties is critical for successfully predicting, re-designing and inhibiting protein-protein interactions. Systematically collected data that link protein sequence to binding are valuable for elucidating determinants of protein interaction, but are rare in the literature because such data are experimentally difficult to generate. Here we describe SORTCERY, a high-throughput method that we have used to rank hundreds of yeast displayed peptides according to their affinities for a target interaction partner. The procedure involves fluorescence-activated cell sorting (FACS) of a library, deep sequencing of sorted pools, and downstream computational analysis. We have developed theoretical models and statistical tools that assist in planning these stages. We demonstrate SORTCERY’s utility by ranking 1026 BH3 peptides with respect to their affinities for the anti-apoptotic protein Bcl-xL. Our results are in striking agreement with measured affinities for 19 individual peptides with dissociation constants ranging from 0.1 to 60 nM. High-resolution ranking can be used to improve our understanding of sequence-function relationships, and to support the development of computational models for predicting and designing novel interactions.

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Section: Introductionmentioning

confidence: 99%

SORTCERY—A High–Throughput Method to Affinity Rank Peptide Ligands

Reich

Dutta

Keating

2015

Journal of Molecular Biology

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“…To develop these radiolabeled peptide probes, we selected a MT1-MMP binding peptide (HWKHLHNTKTFL) that was recently discovered by phage display technology 14) to serve as the core structure of the molecular probe. We then added a Cys residue at the N-or C-terminus of the peptide (LN and LC) to allow facile radiolabeling of the thiol group by N-(m-[ 123/125 I] iodophenyl) maleimide ([ 123/125 I] IPM).…”

mentioning

confidence: 99%

Radioiodinated Peptidic Imaging Probes for <i>in Vivo</i> Detection of Membrane Type-1 Matrix Metalloproteinase in Cancers

Kondo

Temma

Shimizu

et al. 2015

Biological & Pharmaceutical Bulletin

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Membrane type-1 matrix metalloproteinase (MT1-MMP) plays pivotal roles in tumor progression and metastasis, and holds great promise as an early biomarker for malignant tumors. Therefore, the ability to evaluate MT1-MMP expression could be valuable for molecular biological and clinical studies. For this purpose, we aimed to develop short peptide-based nuclear probes because of their facile radiosynthesis, chemically uniform structures, and high specific activity, as compared to antibody-based probes, which could allow them to be more effective for in vivo MT1-MMP imaging. To the best of our knowledge, there have been no reports of radiolabeled peptide probes for the detection of MT1-MMP in cancer tissues. In this study, we designed and prepared four probes which consist of a MT1-MMP-specific binding peptide sequence (consisting of L Key words membrane type-1 matrix metalloproteinase (MT1-MMP); peptide; in vivo imaging Metastasis is the most frequent cause of death in cancer patients.1) In order to metastasize, tumor cells must break through the basement membrane and invade dense networks of the interstitial extracellular matrix (ECM) that surrounds cancer cells.2) Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteases responsible for degrading various ECM components. While most MMPs are secreted as soluble zymogens, a subfamily of membrane-type MMPs (MT-MMPs) is expressed on the cell membrane and mediate pericellular proteolysis.3,4) Among MT-MMPs, MT1-MMP is the most prominent member and is the major pericellular protease involved in degrading triple helical collagen type I.5) In addition, MT1-MMP activates MMP zymogens such as proMMP-2 and proMMP-13, which are also significantly involved in tumor cell invasion and metastasis. 6,7) As MT1-MMP is closely associated with tumor malignancy and holds great promise as an early biomarker of malignant tumors, 8,9) in vivo monitoring of MT1-MMP expression could provide meaningful data for molecular biology and clinical studies.Since nuclear medical techniques are optimal for noninvasive quantitative evaluation of biological molecules deep within the body, we recently developed In-labeled miniaturized antibodies (scFv and diabody ) 11) as radiolabeled probes for nuclear medical imaging of MT1-MMP. Although these probes could be used for imaging MT1-MMP in cancer, antibody derivatives have several undesirable characteristics when used as molecular imaging probes. In addition to the high cost of producing antibody derivatives, 12) radiolabeling of these antibodies could generate heterogeneous mixtures because a radioisotope can non-selectively react with several amino acid residues in a protein sequence depending on the radioisotope introduction reagent. 13) Moreover, radiolabeled high molecular weight probes may show low apparent specific radioactivity (radioactivity per gram) because precursors remaining after the radiolabeling reaction cannot be separated by ordinary size exclusion HPLC methods.11) The presence of substantial amounts of precursor c...

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“…In fact, some fluorogenic probes activated by or binding with MT1-MMP have successfully imaged tumors. (20)(21)(22) However, these probes were insufficient for quantitative analysis of MT1-MMP in vivo. In this regard, radiolabeled probes were expected to achieve more accurate quantitation; therefore, several radiolabeled probes for MT1-MMP have been developed including the 99m Tc-MT1-mAb (9) and 123 I-labelled TIMP-2.…”

Section: Discussionmentioning

confidence: 99%

Miniaturized antibodies for imaging membrane type‐1 matrix metalloproteinase in cancers

et al. 2013

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Since membrane type-1 matrix metalloproteinase (MT1-MMP) plays pivotal roles in tumor progression and metastasis and holds great promise as an early biomarker for malignant tumors, a method of evaluating MT1-MMP expression levels would be valuable for molecular biological and clinical studies. Although we have previously developed a 99m Tc-labeled anti-MT1-MMP monoclonal IgG ( 99m Tc-MT1-mAb) as an MT1-MMP imaging probe by nuclear medical techniques for this purpose, slow pharmacokinetics were a problem due to its large molecular size. Thus, in this study, our aim was to develop miniaturized antibodies, a single chain antibody fragment (MT1-scFv) and a dimer of two molecules of scFv (MT1-diabody), as the basic structures of MT1-MMP imaging probes followed by in vitro and in vivo evaluation with an 111 In radiolabel. Phage display screening successfully provided MT1-scFv and MT1-diabody, which had sufficiently high affinity for MT1-MMP (K D = 29.8 and 17.1 nM). Both 111 In labeled miniaturized antibodies showed higher uptake in MT1-MMP expressing HT1080 cells than in non-expressing MCF7 cells. An in vivo biodistribution study showed rapid pharmacokinetics for both probes, which exhibited >20-fold higher tumor to blood radioactivity ratios (T ⁄ B ratio), an index for in vivo imaging, than 99m Tc-MT1-mAb 6 h post-administration, and significantly higher tumor accumulation in HT1080 than MCF7 cells. SPECT images showed heterogeneous distribution and ex vivo autoradiographic analysis revealed that the radioactivity distribution profiles in tumors corresponded to MT1-MMP-positive areas. These findings suggest that the newly developed miniaturized antibodies are promising probes for detection of MT1-MMP in cancer cells. (Cancer Sci 2013; 104: 495-501) T umor metastasis is the most frequent cause of death for cancer patients. In order to metastasize, tumor cells must acquire the ability to break through the basement membrane and invade dense networks of interstitial ECM proteins.(1) Matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading the various ECM components. While most MMPs are secreted as soluble zymogens, members of the subfamily of membrane-type MMPs (MT-MMPs) anchored to the cell membrane are suited for pericellular proteolysis. MT1-MMP is the major pericellular protease involved in processing triple helical collagen type I.(4) In addition, MT1-MMP activates MMP zymogens such as proMMP-2 and proMMP-13 that have significant involvement in tumor cell invasion and metastasis.(5,6) As MT1-MMP has a close relationship with tumor malignancy and holds great promise as an early biomarker of malignant tumors, (7,8) in vivo monitoring and ⁄ or quantitation of MT1-MMP expression could be valuable tools for molecular biological and clinical studies.Recently, in the course of focusing on nuclear medical techniques for noninvasive quantitative evaluation of biological molecules deep within the body, we developed a 99m Tc-labeled anti-MT1-MMP monoclonal IgG ( 99m Tc-MT1-mAb) as a radiolabeled...

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High-affinity peptide against MT1-MMP for in vivo tumor imaging

Cited by 58 publications

References 41 publications

SORTCERY—A High–Throughput Method to Affinity Rank Peptide Ligands

SORTCERY—A High–Throughput Method to Affinity Rank Peptide Ligands

Radioiodinated Peptidic Imaging Probes for <i>in Vivo</i> Detection of Membrane Type-1 Matrix Metalloproteinase in Cancers

Miniaturized antibodies for imaging membrane type‐1 matrix metalloproteinase in cancers

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