skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Evaluation of integrin αvβ 6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis

Abstract

Advances in precision molecular imaging promise to transform our ability to detect, diagnose and treat disease. Here, we describe the engineering and validation of a new cystine knot peptide (knottin) that selectively recognizes human integrin αvβ 6 with single-digit nanomolar affinity. We solve its 3D structure by NMR and x-ray crystallography and validate leads with 3 different radiolabels in pre-clinical models of cancer. We evaluate the lead tracer's safety, biodistribution and pharmacokinetics in healthy human volunteers, and show its ability to detect multiple cancers (pancreatic, cervical and lung) in patients at two study locations. Additionally, we demonstrate that the knottin PET tracers can also detect fibrotic lung disease in idiopathic pulmonary fibrosis patients. Our results indicate that these cystine knot PET tracers may have potential utility in multiple disease states that are associated with upregulation of integrin αvβ 6.

Authors:
 [1];  [2];  [1];  [2];  [1]; ORCiD logo [3]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [4]; ORCiD logo [1]; ORCiD logo [5];  [6];  [1]; ORCiD logo [1] more »; ORCiD logo [1];  [7];  [1]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [1];  [1];  [2]; ORCiD logo [1] « less
  1. Stanford Univ., CA (United States)
  2. Peking Union Medical College, Beijing (China)
  3. Helmholtz Zentrum München (Germany); Technical Univ. München, Freising (Germany)
  4. Helmholtz Zentrum München (Germany); Technical Univ. München, Freising (Germany); Mississippi State Univ., Mississippi State, MS (United States)
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  6. Art Robbins Instruments, Sunnyvale, CA (United States)
  7. Pliant Therapeutics, Redwood City, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); Canary Foundation; National Institutes of Health (NIH); National Cancer Institute (NCI); GRFP NSF National Science Foundation; Science Alliance on Precision Medicine and Cancer Prevention; National Natural Science Foundation of China (NNSFC); Beijing Municipal Science & Technology Commission; CAMS Innovation Fund for Medical Science
OSTI Identifier:
1580946
Grant/Contract Number:  
AC02-05CH11231; SC0008397; NCI U01 CA210020; CA154887; CRN-17-427258; 81571713; Z151100003915133; 2016-I2M-4-003
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES

Citation Formats

Kimura, Richard H., Wang, Ling, Shen, Bin, Huo, Li, Tummers, Willemieke, Filipp, Fabian V., Guo, Haiwei Henry, Haywood, Thomas, Abou-Elkacem, Lotfi, Baratto, Lucia, Habte, Frezghi, Devulapally, Rammohan, Witney, Timothy H., Cheng, Yan, Tikole, Suhas, Chakraborti, Subhendu, Nix, Jay, Bonagura, Christopher A., Hatami, Negin, Mooney, Joshua J., Desai, Tushar, Turner, Scott, Gaster, Richard S., Otte, Andrea, Visser, Brendan C., Poultsides, George A., Norton, Jeffrey, Park, Walter, Stolowitz, Mark, Lau, Kenneth, Yang, Eric, Natarajan, Arutselvan, Ilovich, Ohad, Srinivas, Shyam, Srinivasan, Ananth, Paulmurugan, Ramasamy, Willmann, Juergen, Chin, Frederick T., Cheng, Zhen, Iagaru, Andrei, Li, Fang, and Gambhir, Sanjiv S. Evaluation of integrin αvβ6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis. United States: N. p., 2019. Web. doi:10.1038/s41467-019-11863-w.
Kimura, Richard H., Wang, Ling, Shen, Bin, Huo, Li, Tummers, Willemieke, Filipp, Fabian V., Guo, Haiwei Henry, Haywood, Thomas, Abou-Elkacem, Lotfi, Baratto, Lucia, Habte, Frezghi, Devulapally, Rammohan, Witney, Timothy H., Cheng, Yan, Tikole, Suhas, Chakraborti, Subhendu, Nix, Jay, Bonagura, Christopher A., Hatami, Negin, Mooney, Joshua J., Desai, Tushar, Turner, Scott, Gaster, Richard S., Otte, Andrea, Visser, Brendan C., Poultsides, George A., Norton, Jeffrey, Park, Walter, Stolowitz, Mark, Lau, Kenneth, Yang, Eric, Natarajan, Arutselvan, Ilovich, Ohad, Srinivas, Shyam, Srinivasan, Ananth, Paulmurugan, Ramasamy, Willmann, Juergen, Chin, Frederick T., Cheng, Zhen, Iagaru, Andrei, Li, Fang, & Gambhir, Sanjiv S. Evaluation of integrin αvβ6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis. United States. doi:10.1038/s41467-019-11863-w.
Kimura, Richard H., Wang, Ling, Shen, Bin, Huo, Li, Tummers, Willemieke, Filipp, Fabian V., Guo, Haiwei Henry, Haywood, Thomas, Abou-Elkacem, Lotfi, Baratto, Lucia, Habte, Frezghi, Devulapally, Rammohan, Witney, Timothy H., Cheng, Yan, Tikole, Suhas, Chakraborti, Subhendu, Nix, Jay, Bonagura, Christopher A., Hatami, Negin, Mooney, Joshua J., Desai, Tushar, Turner, Scott, Gaster, Richard S., Otte, Andrea, Visser, Brendan C., Poultsides, George A., Norton, Jeffrey, Park, Walter, Stolowitz, Mark, Lau, Kenneth, Yang, Eric, Natarajan, Arutselvan, Ilovich, Ohad, Srinivas, Shyam, Srinivasan, Ananth, Paulmurugan, Ramasamy, Willmann, Juergen, Chin, Frederick T., Cheng, Zhen, Iagaru, Andrei, Li, Fang, and Gambhir, Sanjiv S. Mon . "Evaluation of integrin αvβ6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis". United States. doi:10.1038/s41467-019-11863-w. https://www.osti.gov/servlets/purl/1580946.
@article{osti_1580946,
title = {Evaluation of integrin αvβ6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis},
author = {Kimura, Richard H. and Wang, Ling and Shen, Bin and Huo, Li and Tummers, Willemieke and Filipp, Fabian V. and Guo, Haiwei Henry and Haywood, Thomas and Abou-Elkacem, Lotfi and Baratto, Lucia and Habte, Frezghi and Devulapally, Rammohan and Witney, Timothy H. and Cheng, Yan and Tikole, Suhas and Chakraborti, Subhendu and Nix, Jay and Bonagura, Christopher A. and Hatami, Negin and Mooney, Joshua J. and Desai, Tushar and Turner, Scott and Gaster, Richard S. and Otte, Andrea and Visser, Brendan C. and Poultsides, George A. and Norton, Jeffrey and Park, Walter and Stolowitz, Mark and Lau, Kenneth and Yang, Eric and Natarajan, Arutselvan and Ilovich, Ohad and Srinivas, Shyam and Srinivasan, Ananth and Paulmurugan, Ramasamy and Willmann, Juergen and Chin, Frederick T. and Cheng, Zhen and Iagaru, Andrei and Li, Fang and Gambhir, Sanjiv S.},
abstractNote = {Advances in precision molecular imaging promise to transform our ability to detect, diagnose and treat disease. Here, we describe the engineering and validation of a new cystine knot peptide (knottin) that selectively recognizes human integrin αvβ6 with single-digit nanomolar affinity. We solve its 3D structure by NMR and x-ray crystallography and validate leads with 3 different radiolabels in pre-clinical models of cancer. We evaluate the lead tracer's safety, biodistribution and pharmacokinetics in healthy human volunteers, and show its ability to detect multiple cancers (pancreatic, cervical and lung) in patients at two study locations. Additionally, we demonstrate that the knottin PET tracers can also detect fibrotic lung disease in idiopathic pulmonary fibrosis patients. Our results indicate that these cystine knot PET tracers may have potential utility in multiple disease states that are associated with upregulation of integrin αvβ6.},
doi = {10.1038/s41467-019-11863-w},
journal = {Nature Communications},
number = 1,
volume = 10,
place = {United States},
year = {2019},
month = {10}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 1 work
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Targeting of αv integrin identifies a core molecular pathway that regulates fibrosis in several organs
journal, November 2013

  • Henderson, Neil C.; Arnold, Thomas D.; Katamura, Yoshio
  • Nature Medicine, Vol. 19, Issue 12
  • DOI: 10.1038/nm.3282

Roles of αv integrins in vascular biology and pulmonary pathology
journal, October 2004


Comparative Metabolic Flux Profiling of Melanoma Cell Lines: BEYOND THE WARBURG EFFECT
journal, October 2011

  • Scott, David A.; Richardson, Adam D.; Filipp, Fabian V.
  • Journal of Biological Chemistry, Vol. 286, Issue 49
  • DOI: 10.1074/jbc.M111.282046

Defining the Role of Integrin αvβ6 in Cancer
journal, July 2009


Engineered cystine knot peptides that bind αvβ3, αvβ5, and α5β1 integrins with low-nanomolar affinity
journal, November 2009

  • Kimura, Richard H.; Levin, Aron M.; Cochran, Frank V.
  • Proteins: Structure, Function, and Bioinformatics, Vol. 77, Issue 2
  • DOI: 10.1002/prot.22441

Targeted Contrast-Enhanced Ultrasound Imaging of Tumor Angiogenesis with Contrast Microbubbles Conjugated to Integrin-Binding Knottin Peptides
journal, February 2010


New tools to target an elusive integrin
journal, July 2015

  • Kingwell, Katie
  • Nature Reviews Drug Discovery, Vol. 14, Issue 7
  • DOI: 10.1038/nrd4672

Interstitial Lung Disease: NHLBI Workshop on the Primary Prevention of Chronic Lung Diseases
journal, April 2014

  • Rosas, Ivan O.; Dellaripa, Paul F.; Lederer, David J.
  • Annals of the American Thoracic Society, Vol. 11, Issue Supplement 3
  • DOI: 10.1513/AnnalsATS.201312-429LD

The α v β 1 integrin plays a critical in vivo role in tissue fibrosis
journal, May 2015


PHENIX : building new software for automated crystallographic structure determination
journal, October 2002

  • Adams, Paul D.; Grosse-Kunstleve, Ralf W.; Hung, Li-Wei
  • Acta Crystallographica Section D Biological Crystallography, Vol. 58, Issue 11
  • DOI: 10.1107/S0907444902016657

Intraclass correlations: Uses in assessing rater reliability.
journal, January 1979


Subclinical Interstitial Lung Disease: Why You Should Care
journal, June 2012

  • Doyle, Tracy J.; Hunninghake, Gary M.; Rosas, Ivan O.
  • American Journal of Respiratory and Critical Care Medicine, Vol. 185, Issue 11
  • DOI: 10.1164/rccm.201108-1420PP

An Official ATS/ERS/JRS/ALAT Statement: Idiopathic Pulmonary Fibrosis: Evidence-based Guidelines for Diagnosis and Management
journal, March 2011

  • Raghu, Ganesh; Collard, Harold R.; Egan, Jim J.
  • American Journal of Respiratory and Critical Care Medicine, Vol. 183, Issue 6
  • DOI: 10.1164/rccm.2009-040GL

Adenomatous transformation of the human anterior pituitary is associated with alterations in integrin expression
journal, July 1996


Pituitary tumours: findings from whole genome analyses
journal, September 2006


The CCP4 suite programs for protein crystallography
journal, September 1994


XDS
journal, January 2010

  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
  • DOI: 10.1107/S0907444909047337

Partial Inhibition of Integrin αvβ6 Prevents Pulmonary Fibrosis without Exacerbating Inflammation
journal, January 2008

  • Horan, Gerald S.; Wood, Susan; Ona, Victor
  • American Journal of Respiratory and Critical Care Medicine, Vol. 177, Issue 1
  • DOI: 10.1164/rccm.200706-805OC

Use of 64 Cu-labeled Fibronectin Domain with EGFR-Overexpressing Tumor Xenograft: Molecular Imaging
journal, April 2012

  • Hackel, Benjamin J.; Kimura, Richard H.; Gambhir, Sanjiv S.
  • Radiology, Vol. 263, Issue 1
  • DOI: 10.1148/radiol.12111504

Integrin alphavbeta6 mediates the potential for colon cancer cells to colonize in and metastasize to the liver
journal, May 2008


Identification and Characterization of a Peptide with Affinity to Head and Neck Cancer
journal, February 2009

  • Nothelfer, E. -M.; Zitzmann-Kolbe, S.; Garcia-Boy, R.
  • Journal of Nuclear Medicine, Vol. 50, Issue 3
  • DOI: 10.2967/jnumed.108.058123

A Peptide Selected by Biopanning Identifies the Integrin  v 6 as a Prognostic Biomarker for Nonsmall Cell Lung Cancer
journal, June 2007


Integrin–TGF‐β crosstalk in fibrosis, cancer and wound healing
journal, January 2010


The pituitary TGFβ1 system as a novel target for the treatment of resistant prolactinomas
journal, December 2015

  • Recouvreux, M. Victoria; Camilletti, M. Andrea; Rifkin, Daniel B.
  • Journal of Endocrinology, Vol. 228, Issue 3
  • DOI: 10.1530/JOE-15-0451

Integrin-mediated regulation of TGFβ in fibrosis
journal, July 2013

  • Henderson, Neil C.; Sheppard, Dean
  • Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, Vol. 1832, Issue 7
  • DOI: 10.1016/j.bbadis.2012.10.005

Type beta transforming growth factor (TGF-β) is a potent stimulator of the basal secretion of follicle stimulating hormone (FSH) in a pituitary monolayer system
journal, March 1986

  • Ying, Shao-Yao; Becker, Ann; Baird, Andrew
  • Biochemical and Biophysical Research Communications, Vol. 135, Issue 3
  • DOI: 10.1016/0006-291X(86)91020-X

A Cystine Knot Peptide Targeting Integrin  v 6 for Photoacoustic and Fluorescence Imaging of Tumors in Living Subjects
journal, May 2016


Reverse TCA cycle flux through isocitrate dehydrogenases 1 and 2 is required for lipogenesis in hypoxic melanoma cells: Reverse TCA cycle flux in hypoxia by IDH
journal, March 2012


The Effect of Bi-Terminal PEGylation of an Integrin  v 6-Targeted 18F Peptide on Pharmacokinetics and Tumor Uptake
journal, March 2015


Expression of α V-associated integrin β subunits in epithelial ovarian cancer andits relation to prognosis in patients treated with platinum-based regimens
journal, February 2005

  • Maubant, Sylvie; Cruet-Hennequart, Séverine; Dutoit, Soizic
  • Journal of Molecular Histology, Vol. 36, Issue 1-2
  • DOI: 10.1007/s10735-004-4273-0

Preclinical SPECT/CT Imaging of  v 6 Integrins for Molecular Stratification of Idiopathic Pulmonary Fibrosis
journal, October 2013


Thermal, Chemical, and Enzymatic Stability of the Cyclotide Kalata B1:  The Importance of the Cyclic Cystine Knot †
journal, May 2004

  • Colgrave, Michelle L.; Craik, David J.
  • Biochemistry, Vol. 43, Issue 20
  • DOI: 10.1021/bi049711q

αv integrins: key regulators of tissue fibrosis
journal, May 2016

  • Conroy, Kylie P.; Kitto, Laura J.; Henderson, Neil C.
  • Cell and Tissue Research, Vol. 365, Issue 3
  • DOI: 10.1007/s00441-016-2407-9

Selection of optimal molecular targets for tumor-specific imaging in pancreatic ductal adenocarcinoma
journal, May 2017

  • Tummers, Willemieke S.; Farina-Sarasqueta, Arantza; Boonstra, Martin C.
  • Oncotarget, Vol. 8, Issue 34
  • DOI: 10.18632/oncotarget.18232

Pigeon-Breeders' Lung: A Newly Observed Interstitial Pulmonary Disease
journal, July 1965


Integrins in cancer: biological implications and therapeutic opportunities
journal, January 2010

  • Desgrosellier, Jay S.; Cheresh, David A.
  • Nature Reviews Cancer, Vol. 10, Issue 1
  • DOI: 10.1038/nrc2748

NMR View: A computer program for the visualization and analysis of NMR data
journal, September 1994

  • Johnson, Bruce A.; Blevins, Richard A.
  • Journal of Biomolecular NMR, Vol. 4, Issue 5
  • DOI: 10.1007/BF00404272

Cross Talk among TGF-  Signaling Pathways, Integrins, and the Extracellular Matrix
journal, September 2011


Integrin ανβ6 Acts as a Prognostic Indicator in Gastric Carcinoma
journal, February 2008


A Mechanism for Regulating Pulmonary Inflammation and Fibrosis: The Integrin αvβ6 Binds and Activates Latent TGF β1
journal, February 1999


αvβ6 integrin may be a potential prognostic biomarker in interstitial lung disease
journal, March 2015

  • Saini, Gauri; Porte, Joanne; Weinreb, Paul H.
  • European Respiratory Journal, Vol. 46, Issue 2
  • DOI: 10.1183/09031936.00210414

Immunohistochemical demonstration of TGF-beta-receptor type II in human pituitary adenomas
journal, January 1995


Epidemiology of idiopathic pulmonary fibrosis
journal, November 2013


Identification of a Novel ITGα v β 6 -Binding Peptide Using Protein Separation and Phage Display
journal, May 2017


Automated evaluation of chemical shift perturbation spectra: New approaches to quantitative analysis of receptor-ligand interaction NMR spectra
journal, August 2004


Conformational Plasticity of the Lipid Transfer Protein SCP2 †
journal, July 2007

  • Filipp, Fabian V.; Sattler, Michael
  • Biochemistry, Vol. 46, Issue 27
  • DOI: 10.1021/bi6025616

Probing lipid- and drug-binding domains with fluorescent dyes
journal, February 2008

  • Black, Shannon L.; Stanley, Will A.; Filipp, Fabian V.
  • Bioorganic & Medicinal Chemistry, Vol. 16, Issue 3
  • DOI: 10.1016/j.bmc.2007.10.080

NMRPipe: A multidimensional spectral processing system based on UNIX pipes
journal, November 1995

  • Delaglio, Frank; Grzesiek, Stephan; Vuister, GeertenW.
  • Journal of Biomolecular NMR, Vol. 6, Issue 3
  • DOI: 10.1007/BF00197809

Integrins
journal, September 2002


Topography for Independent Binding of α-Helical and PPII-Helical Ligands to a Peroxisomal SH3 Domain
journal, November 2002


Engineered Knottin Peptides: A New Class of Agents for Imaging Integrin Expression in Living Subjects
journal, March 2009


Overexpression of the αvβ6 integrin in cervical squamous cell carcinoma is a prognostic factor for decreased survival
journal, January 2007

  • Hazelbag, S.; Kenter, Gg; Gorter, A.
  • The Journal of Pathology, Vol. 212, Issue 3
  • DOI: 10.1002/path.2168

Rgd and Other Recognition Sequences for Integrins
journal, November 1996


Transcriptional activation of integrin β6 during the epithelial-mesenchymal transition defines a novel prognostic indicator of aggressive colon carcinoma
journal, January 2005

  • Bates, Richard C.; Bellovin, David I.; Brown, Courtney
  • Journal of Clinical Investigation, Vol. 115, Issue 2
  • DOI: 10.1172/JCI200523183

High numbers of human skin cancers express MMP2 and several integrin genes
journal, March 2008


Integrin αvβ6 is a marker of the progression of biliary and portal liver fibrosis and a novel target for antifibrotic therapies
journal, March 2008


Radiolabelled RGD peptides for imaging and therapy
journal, February 2012

  • Gaertner, F. C.; Kessler, H.; Wester, H. -J.
  • European Journal of Nuclear Medicine and Molecular Imaging, Vol. 39, Issue S1
  • DOI: 10.1007/s00259-011-2028-1

Definition of an Unexpected Ligand Recognition Motif for αvβ6 Integrin
journal, January 1999

  • Kraft, Sabine; Diefenbach, Beate; Mehta, Raj
  • Journal of Biological Chemistry, Vol. 274, Issue 4
  • DOI: 10.1074/jbc.274.4.1979

Recognition of a Functional Peroxisome Type 1 Target by the Dynamic Import Receptor Pex5p
journal, December 2006


Labeling strategies for 13C-detected aligned-sample solid-state NMR of proteins
journal, December 2009

  • Filipp, Fabian V.; Sinha, Neeraj; Jairam, Lena
  • Journal of Magnetic Resonance, Vol. 201, Issue 2
  • DOI: 10.1016/j.jmr.2009.08.012

Clinical Application of Radiolabeled RGD Peptides for PET Imaging of Integrin α v β 3
journal, January 2016


First Experience with Clinical-Grade [18F]FPP(RGD)2: An Automated Multi-step Radiosynthesis for Clinical PET Studies
journal, March 2011

  • Chin, Frederick T.; Shen, Bin; Liu, Shuanglong
  • Molecular Imaging and Biology, Vol. 14, Issue 1
  • DOI: 10.1007/s11307-011-0477-3

Phaser crystallographic software
journal, July 2007

  • McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
  • Journal of Applied Crystallography, Vol. 40, Issue 4
  • DOI: 10.1107/S0021889807021206