To map the cellular topography of the rare 3-O-sulfated structural motif of heparan sulfate (HS), we constructed quantum dot-based probes for antithrombin and FGF2, which reveal widely different distribution of the targeted HS motifs. The technology helps show that old and young aortic endothelia display widely different levels of the antithrombin-binding 3-O-sulfated HS motif.

Curcumin and trans- cinnamaldehyde are acrolein-based Michael acceptor compounds that are commonly found in domestic condiments, and known to cause cancer cell death via redox mechanisms. Based on the structural features of these compounds we designed and synthesized several 2-cinnamamido-N-substituted-cinnamamide (bis-cinnamamide) compounds. One of the derivatives, ( Z )-2-[( E )-cinnamamido]-3-phenyl-N-propylacrylamide ( 1512) showed a moderate antiproliferative potency (HT116 cell line inhibition of 32.0 µM ± 2.6) with proven cellular activities leading to apoptosis. Importantly, 1512 exhibited good selectivity toxicity on cancer cells over noncancerous cells (IC ₅₀ of C-166 cell lines >100 µM), and low cancer cell resistance at 100 µM dose (growth rate 10.1±1.1%). We subsequently carried out structure activity relationship studies with 1512 . Derivatives with electron rich moiety at the aryl ring of the 2-aminocinnamaide moiety exhibited strong antiproliferative action while electron withdrawing groups caused loss of activity. Our most promising compound, 4112 [( Z )-3-(1 H -indol-3-yl)- N -propyl-2-[( E )-3-(thien-2-yl)propenamido)propenamide] killed cancer cells at IC ₅₀ = 0.89 ± 0.04 µM (Caco-2), 2.85 ± 1.5 (HCT-116) and 1.65 ± 0.07 (HT-29), while exhibiting much weaker potency on C-166 and BHK normal cell lines (IC ₅₀ = 71 ± 5.12 and 77.6 ± 6.2 µM, respectively). Cellular studies towards identifying the compounds mechanism of cytotoxic activities revealed that apoptotic induction occurs in part due to oxidative stress. Importantly, the compounds showed inhibition of cancer stem cells that are critical for maintaining the potential for self-renewal and stemness. The results presented here show discovery of Michael addition compounds that potently kill cancer cells by a defined mechanism, with minimal effect on normal noncancerous cell.

Figure S2. StarD5 protein is increased in most human colorectal tumors compared to adjacent normal tissue.

Figure S10. PM cholesterol levels in Dual hi CRCSC.

Figure S4. StarD5 protein expression by immunofluorescence (IF) compared to adjacent normal tissue.

Figure S9. PM cholesterol levels in CRCSC.