Effective Inhibition of Bone Morphogenetic Protein Function by Highly Specific Llama-Derived Antibodies.
Bone morphogenetic proteins (BMP) have important but distinct roles in tissue homeostasis and disease, including carcinogenesis and tumor progression. A large number of BMP inhibitors are available to study BMP function; however, as most of these antagonists are promiscuous, evaluating specific effects of individual BMPs is not feasible. Because the oncogenic role of the different BMPs varies for each neoplasm, highly selective BMP inhibitors are required. Here, we describe the generation of three types of llama-derived heavy chain variable domains (VHH) that selectively bind to either BMP4, to BMP2 and 4, or to BMP2, 4, 5, and 6. These generated VHHs have high affinity to their targets and are able to inhibit BMP signaling. Epitope binning and docking modeling have shed light into the basis for their BMP specificity. As opposed to the wide structural reach of natural inhibitors, these small molecules target the grooves and pockets of BMPs involved in receptor binding. In organoid experiments, specific inhibition of BMP4 does not affect the activation of normal stem cells. Furthermore, in vitro inhibition of cancer-derived BMP4 noncanonical signals results in an increase of chemosensitivity in a colorectal cancer cell line. Therefore, because of their high specificity and low off-target effects, these VHHs could represent a therapeutic alternative for BMP4(+) malignancies.
Center for Experimental and Molecular Medicine, Academic Medical Center, Amsterdam, the Netherlands.
Silvia
S
Calpe
AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands.
Koen
K
Wagner
QVQ BV, Utrecht, the Netherlands.
Mohamed
M
El Khattabi
QVQ BV, Utrecht, the Netherlands.
Lucy
L
Rutten
Laboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
Cheryl
C
Zimberlin
QVQ BV, Utrecht, the Netherlands.
Edward
E
Dolk
QVQ BV, Utrecht, the Netherlands.
C Theo
CT
Verrips
Laboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
Jan Paul
JP
Medema
AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands.
Hergen
H
Spits
Center for Experimental and Molecular Medicine, Academic Medical Center, Amsterdam, the Netherlands. Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, the Netherlands. k.k.krishnadath@amc.uva.nl.
Kausilia K
KK
Krishnadath
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D002161:Camelids, New World / Q000276:immunology*; D002460:Cell Line; D019073:HT29 Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009369:Neoplasms / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary
D000906:Antibodies; C525718:BMP2 protein, human; C525809:BMP4 protein, human; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins