Cholinergic signaling via muscarinic M1 receptor confers resistance to docetaxel in prostate cancer.
Docetaxel is the most commonly used chemotherapy for advanced prostate cancer (PC), including castration-resistant disease (CRPC), but the eventual development of docetaxel resistance constitutes a major clinical challenge. Here, we demonstrate activation of the cholinergic muscarinic M1 receptor (CHRM1) in CRPC cells upon acquiring resistance to docetaxel, which is manifested in tumor tissues from PC patients post- vs. pre-docetaxel. Genetic and pharmacological inactivation of CHRM1 restores the efficacy of docetaxel in resistant cells. Mechanistically, CHRM1, via its first and third extracellular loops, interacts with the SEMA domain of cMET and forms a heteroreceptor complex with cMET, stimulating a downstream mitogen-activated protein polykinase program to confer docetaxel resistance. Dicyclomine, a clinically available CHRM1-selective antagonist, reverts resistance and restricts the growth of multiple docetaxel-resistant CRPC cell lines and patient-derived xenografts. Our study reveals a CHRM1-dictated mechanism for docetaxel resistance and identifies a CHRM1-targeted combinatorial strategy for overcoming docetaxel resistance in PC.
Pubmed ID: 38262412 RIS Download
Research resources used in this publication
- pCMV-VSV-G (RRID:Addgene_8454) (plasmid)
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NOD.Cg-Prkdcscid Il2rg
- J:NU (RRID:IMSR_JAX:007850) (organism)
- WPMY-1 (RRID:CVCL_3814) (cell line)
- NCI-H660 (RRID:CVCL_1576) (cell line)
- LASCPC-01 (RRID:CVCL_UE17) (cell line)
- PC-3 (RRID:CVCL_0035) (cell line)
- COS-1 (RRID:CVCL_0223) (cell line)
- HEK293T (RRID:CVCL_0063) (cell line)
- DU145 (RRID:CVCL_0105) (cell line)
- 22Rv1 (RRID:CVCL_1045) (cell line)
- pCMV delta R8.2 (RRID:Addgene_12263) (plasmid)
- pCDNA3-Flag MKK6(glu) (RRID:Addgene_13518) (plasmid)
- pCMV-myc-ERK2-L4A-MEK1_fusion (RRID:Addgene_39197) (plasmid)
- FLAG-Mkk7-JNK2 fusion-pcw107 (RRID:Addgene_64618) (plasmid)
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Antibodies used in this publication
- Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 555 (RRID:AB_2633281)
- Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (RRID:AB_2633275)
- mouse anti-goat IgG-HRP (RRID:AB_628490)
- Anti-rabbit IgG, HRP-linked Antibody (RRID:AB_2099233)
- goat anti-mouse IgG-HRP (RRID:AB_631737)
- p63 (D-9) (RRID:AB_628092)
- VENTANA anti-p63 (4A4) Mouse Monoclonal Primary Antibody (RRID:AB_2335989)
- P504S (RRID:AB_2305454)
- Beta Arrestin 2 antibody (RRID:AB_10644158)
- beta-Actin (AC-15) (RRID:AB_1119529)
- Anti-VAChT (RRID:AB_887864)
- Monoclonal ANTI-FLAG® M2 antibody produced in mouse (RRID:AB_262044)
- SYP (D-4) (RRID:AB_628311)
- PKCdelta (D10E2) Rabbit mAb (RRID:AB_10949973)
- PKC Alpha antibody (RRID:AB_2878965)
- Phospho-PKCdelta (Tyr311) Antibody (RRID:AB_330876)
- Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP(tm) Rabbit mAb (RRID:AB_2139682)
- Phospho-PKC Alpha (Thr638) Antibody (RRID:AB_2918239)
- Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb (RRID:AB_823588)
- p-ERK (Tyr 204) (RRID:AB_2297323)
- p38 MAPK (D13E1) XP Rabbit mAb (RRID:AB_10999090)
- Rabbit Anti-Met, phospho (Tyr1234 / Tyr1235) Monoclonal Antibody, Unconjugated, Clone D26 (RRID:AB_2143884)
- Normal Rabbit IgG (RRID:AB_1031062)
- NF-L antibody (RRID:AB_10860271)
- Ki-67 (D2H10) Rabbit mAb (IHC Specific) (RRID:AB_2636984)
- SAPK/JNK Antibody (RRID:AB_2250373)
- Galpha q/11/14 (G-7) (RRID:AB_10842057)
- ERK 1/2 Antibody (C-9) (RRID:AB_2571739)
- Met (D1C2) XP Rabbit mAb (RRID:AB_10858224)
- Cleaved PARP (Asp214) Antibody (Human Specific) (RRID:AB_331426)
- Met (L6E7) Mouse mAb (RRID:AB_11127596)
- Cleaved Caspase-3 (Asp175) Antibody (RRID:AB_2341188)
- Anti-CHRM1 antibody produced in rabbit (RRID:AB_1854210)
- Anti-Choline Acetyltransferase antibody [EPR13024(B)] (RRID:AB_2687983)
- Acetylcholinesterase antibody (RRID:AB_2242349)
Associated grants
- Agency: NCI NIH HHS, United States
Id: R01 CA258634 - Agency: NCI NIH HHS, United States
Id: R37 CA233658
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View all literature mentionsGraphPad Prism (tool)
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Software package for interpreting gene expression data. Used for interpretation of a large-scale experiment by identifying pathways and processes.
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Software ultrafast memory efficient tool for aligning sequencing reads. Bowtie is short read aligner.
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RRID:AB_631737
This polyclonal targets goat anti-mouse IgG-HRP
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This monoclonal targets p63
View all literature mentionsBeta Arrestin 2 antibody (antibody)
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This polyclonal targets Beta Arrestin 2
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This monoclonal targets beta-Actin (AC-15)
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This polyclonal targets PKC Alpha
View all literature mentionsPhospho-PKCdelta (Tyr311) Antibody (antibody)
RRID:AB_330876
This polyclonal targets Phospho-PKCdelta (Tyr311)
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This monoclonal targets Mapk14
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RRID:AB_2297323
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View all literature mentionsp38 MAPK (D13E1) XP Rabbit mAb (antibody)
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This polyclonal targets Cleaved Caspase-3 (Asp175)
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RRID:AB_1854210
This unknown targets Human CHRM1
View all literature mentionsAnti-Choline Acetyltransferase antibody [EPR13024(B)] (antibody)
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This monoclonal targets Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Choline Acetyltransferase aa 700 to the C-terminus. The exact sequence is proprietary.
View all literature mentionsAcetylcholinesterase antibody (antibody)
RRID:AB_2242349
This polyclonal targets Acetylcholinesterase
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NOD.Cg-Prkdcscid Il2rg/SzJ (organism)
RRID:IMSR_JAX:005557
Mus musculus with name NOD.Cg-Prkdcscid Il2rg
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Mus musculus with name J:NU from IMSR.
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Cell line WPMY-1 is a Transformed cell line with a species of origin Homo sapiens
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Cell line NCI-H660 is a Cancer cell line with a species of origin Homo sapiens (Human)
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Cell line LASCPC-01 is a Cancer cell line with a species of origin Homo sapiens
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RRID:CVCL_0223
Cell line COS-1 is a Transformed cell line with a species of origin Chlorocebus aethiops (Green monkey)
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RRID:CVCL_0063
Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentionsDU145 (cell line)
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Cell line DU145 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentions22Rv1 (cell line)
RRID:CVCL_1045
Cell line 22Rv1 is a Cancer cell line with a species of origin Homo sapiens (Human)
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