PMID- 34948407
OWN - NLM
STAT- MEDLINE
DCOM- 20220121
LR  - 20220512
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 22
IP  - 24
DP  - 2021 Dec 19
TI  - Single-Dose P2 X4R Single-Chain Fragment Variable Antibody Permanently Reverses 
      Chronic Pain in Male Mice.
LID - 10.3390/ijms222413612 [doi]
LID - 13612
AB  - Non-opioid single-chain variable fragment (scFv) small antibodies were generated 
      as pain-reducing block of P2X4R receptor (P2X4R). A panel of scFvs targeting an 
      extracellular peptide sequence of P2X4R was generated followed by cell-free 
      ribosome display for recombinant antibody selection. After three rounds of 
      bio-panning, a panel of recombinant antibodies was isolated and characterized by 
      ELISA, cross-reactivity analysis, and immunoblotting/immunostaining. Generated 
      scFv antibodies feature binding activity similar to monoclonal antibodies but 
      with stronger affinity and increased tissue penetrability due to their ~30% 
      smaller size. Two anti-P2X4R scFv clones (95, 12) with high specificity and 
      affinity binding were selected for in vivo testing in male and female mice with 
      trigeminal nerve chronic neuropathic pain (FRICT-ION model) persisting for 
      several months in untreated BALBc mice. A single dose of P2X4R scFv (4 mg/kg, 
      i.p.) successfully, completely, and permanently reversed chronic neuropathic 
      pain-like measures in male mice only, providing retention of baseline behaviors 
      indefinitely. Untreated mice retained hypersensitivity, and developed anxiety- 
      and depression-like behaviors within 5 weeks. In vitro P2X4R scFv 95 treatment 
      significantly increased the rheobase of larger-diameter (>25 microm) trigeminal 
      ganglia (TG) neurons from FRICT-ION mice compared to controls. The data support 
      use of engineered scFv antibodies as non-opioid biotherapeutic interventions for 
      chronic pain.
FAU - Westlund, Karin N
AU  - Westlund KN
AUID- ORCID: 0000-0002-7388-4834
AD  - Department of Anesthesiology & Critical Care Medicine, University of New Mexico 
      Health Sciences Center, Albuquerque, NM 87131, USA.
AD  - Biomedical Laboratory Research & Development (121F), New Mexico VA Health Care 
      System, Albuquerque, NM 87108, USA.
FAU - Montera, Marena A
AU  - Montera MA
AD  - Department of Anesthesiology & Critical Care Medicine, University of New Mexico 
      Health Sciences Center, Albuquerque, NM 87131, USA.
FAU - Goins, Aleyah E
AU  - Goins AE
AD  - Department of Anesthesiology & Critical Care Medicine, University of New Mexico 
      Health Sciences Center, Albuquerque, NM 87131, USA.
FAU - Alles, Sascha R A
AU  - Alles SRA
AD  - Department of Anesthesiology & Critical Care Medicine, University of New Mexico 
      Health Sciences Center, Albuquerque, NM 87131, USA.
FAU - Suri, Nikita
AU  - Suri N
AD  - Ross University Medical Center, Miramar, FL 60153, USA.
FAU - McIlwrath, Sabrina L
AU  - McIlwrath SL
AD  - Biomedical Laboratory Research & Development (121F), New Mexico VA Health Care 
      System, Albuquerque, NM 87108, USA.
FAU - Bartel, Robyn
AU  - Bartel R
AD  - Department of Anesthesiology & Critical Care Medicine, University of New Mexico 
      Health Sciences Center, Albuquerque, NM 87131, USA.
FAU - Durvasula, Ravi V
AU  - Durvasula RV
AD  - Department of Medicine, Mayo Clinic, Jacksonville, FL 32224, USA.
FAU - Kunamneni, Adinarayana
AU  - Kunamneni A
AUID- ORCID: 0000-0002-9952-2209
AD  - Department of Medicine, Mayo Clinic, Jacksonville, FL 32224, USA.
LA  - eng
GR  - I01 BX002695/BX/BLRD VA/United States
GR  - R21 DE028096/DE/NIDCR NIH HHS/United States
GR  - UG3 NS123958/NS/NINDS NIH HHS/United States
GR  - W81XWH2010930/United States Department of Defense/
PT  - Journal Article
DEP - 20211219
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Peptide Library)
RN  - 0 (Purinergic P2X Receptor Antagonists)
RN  - 0 (Receptors, Purinergic P2X4)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Single-Chain Antibodies)
SB  - IM
MH  - Animals
MH  - Antibody Affinity
MH  - Cells, Cultured
MH  - Chronic Pain/*drug therapy/immunology
MH  - Female
MH  - Male
MH  - Mice
MH  - Peptide Library
MH  - Purinergic P2X Receptor Antagonists/chemistry/immunology/*therapeutic use
MH  - Receptors, Purinergic P2X4/chemistry/immunology
MH  - Recombinant Proteins/chemistry/immunology/therapeutic use
MH  - Single-Chain Antibodies/chemistry/immunology/*therapeutic use
PMC - PMC8706307
OTO - NOTNLM
OT  - anxiety
OT  - chronic pain
OT  - depression
OT  - hypersensitivity
OT  - nerve injury
OT  - neuropathy
OT  - pain therapy
OT  - small antibody
COIS- The authors declare no conflict of interest.
EDAT- 2021/12/25 06:00
MHDA- 2022/01/22 06:00
PMCR- 2021/12/19
CRDT- 2021/12/24 01:17
PHST- 2021/10/26 00:00 [received]
PHST- 2021/12/02 00:00 [revised]
PHST- 2021/12/16 00:00 [accepted]
PHST- 2021/12/24 01:17 [entrez]
PHST- 2021/12/25 06:00 [pubmed]
PHST- 2022/01/22 06:00 [medline]
PHST- 2021/12/19 00:00 [pmc-release]
AID - ijms222413612 [pii]
AID - ijms-22-13612 [pii]
AID - 10.3390/ijms222413612 [doi]
PST - epublish
SO  - Int J Mol Sci. 2021 Dec 19;22(24):13612. doi: 10.3390/ijms222413612.