PMID- 36343233
OWN - NLM
STAT- MEDLINE
DCOM- 20221109
LR  - 20221121
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 119
IP  - 46
DP  - 2022 Nov 16
TI  - Directed evolution identifies high-affinity cystine-knot peptide agonists and 
      antagonists of Wnt/beta-catenin signaling.
PG  - e2207327119
LID - 10.1073/pnas.2207327119 [doi]
LID - e2207327119
AB  - Developing peptide-based tools to fine-tune growth signaling pathways, in 
      particular molecules with exquisite selectivity and high affinities, opens up 
      opportunities for cellular reprogramming in tissue regeneration. Here, we present 
      a library based on cystine-knot peptides (CKPs) that incorporate multiple loops 
      for randomization and selection via directed evolution. Resulting binders could 
      be assembled into multimeric structures to fine-tune cellular signaling. An 
      example is presented for the Wnt pathway, which plays a key role in the 
      homeostasis and regeneration of tissues such as lung, skin, and intestine. We 
      discovered picomolar affinity CKP agonists of the human LPR6 receptor by 
      exploring the limits of the topological manipulation of LRP6 dimerization. 
      Structural analyses revealed that the agonists bind at the first beta-propeller 
      domain of LRP6, mimicking the natural Wnt inhibitors DKK1 and SOST. However, the 
      CKP agonists exhibit a different mode of action as they amplify the signaling of 
      natural Wnt ligands but do not activate the pathway by themselves. In an 
      alveolosphere organoid model, the CKP agonists induced alveolar stem cell 
      activity. They also stimulated growth in primary human intestinal organoids. The 
      approach described here advances the important frontier of next-generation 
      agonist design and could be applied to other signaling pathways to discover 
      tunable agonist ligands.
FAU - Hansen, Simon
AU  - Hansen S
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Zhang, Yingnan
AU  - Zhang Y
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Hwang, Sunhee
AU  - Hwang S
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Nabhan, Ahmad
AU  - Nabhan A
AD  - Department of Physiological Chemistry, Genentech, South San Francisco, CA.
FAU - Li, Wanqing
AU  - Li W
AUID- ORCID: 0000-0001-6432-5195
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Fuhrmann, Jakob
AU  - Fuhrmann J
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Kschonsak, Yvonne
AU  - Kschonsak Y
AUID- ORCID: 0000-0002-8057-2564
AD  - Department of Molecular Oncology, Genentech, South San Francisco, CA.
FAU - Zhou, Lijuan
AU  - Zhou L
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Nile, Aaron H
AU  - Nile AH
AUID- ORCID: 0000-0002-7370-8769
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Gao, Xinxin
AU  - Gao X
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
FAU - Piskol, Robert
AU  - Piskol R
AD  - Department of Molecular Oncology, Genentech, South San Francisco, CA.
FAU - de Sousa E Melo, Felipe
AU  - de Sousa E Melo F
AD  - Department of Molecular Oncology, Genentech, South San Francisco, CA.
FAU - de Sauvage, Frederic J
AU  - de Sauvage FJ
AUID- ORCID: 0000-0002-5275-2584
AD  - Department of Molecular Oncology, Genentech, South San Francisco, CA.
FAU - Hannoush, Rami N
AU  - Hannoush RN
AD  - Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA.
LA  - eng
PT  - Journal Article
DEP - 20221107
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (beta Catenin)
RN  - 0 (Low Density Lipoprotein Receptor-Related Protein-6)
RN  - 0 (Wnt Proteins)
RN  - 48TCX9A1VT (Cystine)
RN  - 0 (Ligands)
RN  - 0 (Peptides)
SB  - IM
MH  - Humans
MH  - *Wnt Signaling Pathway
MH  - *beta Catenin/metabolism
MH  - Low Density Lipoprotein Receptor-Related Protein-6/genetics/metabolism
MH  - Wnt Proteins/metabolism
MH  - Cystine
MH  - Ligands
MH  - Peptides
PMC - PMC9674260
OTO - NOTNLM
OT  - Wnt
OT  - agonist
OT  - directed evolution
OT  - peptide
OT  - phage display
OT  - regeneration
OT  - stem cells
COIS- Competing interest statement: All of the authors were employees of Genentech, a 
      member of the Roche group, at the time the research was performed.
EDAT- 2022/11/08 06:00
MHDA- 2022/11/10 06:00
PMCR- 2022/11/07
CRDT- 2022/11/07 15:23
PHST- 2022/11/07 15:23 [entrez]
PHST- 2022/11/08 06:00 [pubmed]
PHST- 2022/11/10 06:00 [medline]
PHST- 2022/11/07 00:00 [pmc-release]
AID - 202207327 [pii]
AID - 10.1073/pnas.2207327119 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2022 Nov 16;119(46):e2207327119. doi: 
      10.1073/pnas.2207327119. Epub 2022 Nov 7.