Novel regulatory network of the TGF-β/ZEB1/LMO7 axis in colorectal cancer metastasis

Table 1 Knowledge gaps and future research directions for LIM domain only protein 7 in colorectal cancer

Research area	Unresolved issues	Suggested approach
Clinical expression and prognosis	Lack of large-scale, multi-center, independent cohort validation	Conduct a multi-center retrospective study; perform single-cell sequencing to analyze the expression heterogeneity of LMO7 in cell subpopulations
Upstream regulatory mechanism	Does ZEB1 regulate LMO7 through epigenetic modification?	Perform ChIP-qPCR for histone modifications at the LMO7 promoter; validate using HDAC inhibitor intervention
Cellular function and molecular mechanism	Does LMO7 affect the membrane localization of E-cadherin? Downstream interacting proteins are unknown	Use immunofluorescence to observe membrane localization and Co-IP-MS to identify interacting proteins; generate LIM domain-deletion mutants to define its function
Subcellular localization and function	What triggers LMO7 nuclear translocation? What are its nuclear functions?	Perform nuclear-cytoplasmic fractionation combined with immunofluorescence and DNA pull-down-MS to identify nuclear binding complexes

Co-IP-MS: Co-immunoprecipitation mass spectrometry; ChIP-seq: Chromatin immunoprecipitation sequencing; LMO7: LIM domain only protein 7; ZEB1: Zinc finger E-box binding homeobox 1; ChIP-qPCR: Chromatin immunoprecipitation quantitative polymerase chain reaction; HDAC: Histone deacetylase.