High-throughput Screening targets neurotransmitter signaling in small cell lung carcinoma.

Improgo MR, Johnson CW, Tapper AR, Gardner PD. "PLoS One. 2011"

Intruduction:

A nuclear magnetic resonance-based ligand screening method making use of a paramagnetic lanthanide probe is offered. By repairing a paramagnetic lanthanide ion to a goal protein, a pseudo-contact shift (PCS) plus a paramagnetic relaxation enhancement (PRE) could be noticed for both the goal protein and its bound ligand. According to PRE and PCS data, the bound ligand is then screened from the compound library as well as the construction with the ligand-protein complicated is determined.

PRE is definitely an isotropic paramagnetic impact noticed inside 30 Å in the lanthanide ion, and it is utilized for the ligand screening inside the present research. PCS is an anisotropic paramagnetic effect providing long-range (~40 Å) distance and angular info on the noticed nuclei relative to the paramagnetic lanthanide ion, and used for the construction perseverance from the ligand-protein complex.

Since a two-point anchored lanthanide-binding peptide tag is used for repairing the lanthanide ion for the target protein, this screening strategy might be typically applied to non-metal-binding proteins. The usefulness of this technique was demonstrated in the situation of the growth element receptor-bound protein 2 (Grb2) Src homology two (SH2) domain and its low- and high-affinity ligands.

BACKGROUND:

Frontline treatment of small cell lung carcinoma (SCLC) relies greatly on chemotherapeutic agents and radiation treatment. Although SCLC individuals reply properly to first cycles of chemotherapy, they eventually develop resistance. Identification of novel therapies towards SCLC is as a result imperative.

METHODS AND FINDINGS:

We have created a bioluminescence-based cell viability assay for high-throughput screening of anti-SCLC agents. The assay was very first validated via common pharmacological agents and RNA interference employing two human SCLC cell lines. We then utilized the assay in a high-throughput screen employing the LOPAC(1280) compound library. The screening identified several drugs that target traditional most cancers signaling pathways also as neuroendocrine markers in SCLC. In specific, perturbation of dopaminergic and serotonergic signaling inhibits SCLC cell viability.

CONCLUSIONS:

The convergence of our pharmacological information with important SCLC pathway parts reiterates the importance of neurotransmitter signaling in SCLC etiology and points to possible qualified prospects for drug improvement.