Identification of 2-[4-[(4-Methoxyphenyl)methoxy]-phenyl]acetonitrile and Derivatives as Potent Oct3/4 Inducers

Abstract

Reprogramming somatic cells into induced-pluripotent cells (iPSCs) provides new access to all somatic cell types for clinical application without any ethical controversy arising from the use of embryonic stem cells (ESCs). Established protocols for iPSCs generation based on viral transduction with defined factors are limited by low efficiency and the risk of genetic abnormality. Several small molecules have been reported as replacements for defined transcriptional factors, but a chemical able to replace Oct3/4 allowing the generation of human iPSCs is still unavailable. Using a cell-based High Throughput Screening (HTS) campaign, we identified that 2-[4-[(4-methoxyphenyl)methoxy]phenyl]acetonitrile (1), termed O4I1, enhanced Oct3/4 expression. Structural verification and modification by chemical synthesis showed that O4I1 and its derivatives not only promoted expression and stabilization of Oct3/4 but also enhanced its transcriptional activity in diverse human somatic cells, implying the possible benefit from using this class of compounds in regenerative medicine.

The essential role of TAp73 in bortezomib-induced apoptosis in p53-deficient colorectal cancer cells

“…As we described before 49, 50 , HCT116 cells were seeded in a Ø-12mm cover slip coated with Geltrex (Life Technologies, Germany) at a density of 200,000 cells/well. After 24 hrs the cells were incubated with bortezomib at designed concentrations for 24 hrs, fixed with 4% PFA at RT (room temperature) for 15 min, and blocked with blocking buffer (5% goat serum, 1% BSA and 0.3% Triton X-100 in PBS) for 30 min.…”

“…The cells were trypsinized, resuspended in PBS, fixed with 4% PFA and blocked in blocking buffer for 1 hrs 49 . The suspension was incubated with either cleaved PARP or caspase 3 over night.…”

Methylisoindigo preferentially kills cancer stem cells by interfering cell metabolism via inhibition of LKB1 and activation of AMPK in PDACs

“…Meisoindigo represses activity of CSC-associated genes and reduces stemness of Jopaca-1 A body of experimental results indicates that genes involved in the regulation of pluripotency, such as Oct3/4, Nanog and Sox2 (Cheng et al, 2015a(Cheng et al, , 2015bLonardo et al, 2011), or abundantly expressed during EMT (Chaffer et al, 2013), such as N-Cadherin and ZEB1, could play important roles in maintaining CSCs. Upon treatment with 20 mM meisoindigo for 24 h, we detected a significant reduction in the expression of CSCassociated genes in Jopaca-1 analyzed by qRT-PCR, showing only 40% residual expression for ZEB1 and 10% for Oct3/4, Sox2 and N-Cadherin (Figure 2A).…”

Methylisoindigo and Its Bromo-Derivatives Are Selective Tyrosine Kinase Inhibitors, Repressing Cellular Stat3 Activity, and Target CD133+ Cancer Stem Cells in PDAC

“…Human primary fibroblasts were isolated and cultivated as described previously with positive ethic permission [ 59 , 60 ].…”

“…As previously reported [ 18 , 59 , 60 ], 2 × 10 5 cells were seeded and treated with compounds for 30 min or 2 h. Cells were lysed in Urea-lysis buffer containing 1 mM EDTA, 0.5% Triton X-100, 5 mM NaF, 6 M Urea, 1 mM Na 3 VO 4 , 10 mg/mL Pepstatin, 100 mM PMSF, and 3 mg/mL Aprotinin in PBS. Protein concentrations were normalized to the smallest value and proteins were resolved on 8% SDS-PAGE and blotted to membrane using BlueFlash Large semi-dry blotter from Serva Electrophoresis (Heidelberg, Germany).…”