Targeting endogenous proteins for degradation through the affinity-directed protein missile system.

Targeted proteolysis of endogenous proteins is desirable as a research toolkit and in therapeutics. CRISPR/Cas9-mediated gene knockouts are irreversible and often not feasible for many genes. Similarly, RNA interference approaches necessitate prolonged treatments, can lead to incomplete knockdowns and are often associated with off-target effects. Targeted proteolysis can overcome these limitations. In this report, we describe an affinity-directed protein missile (AdPROM) system that harbours the von Hippel-Lindau (VHL) protein, the substrate receptor of the Cullin2 (CUL2) E3 ligase complex, tethered to polypeptide binders that selectively bind and recruit endogenous target proteins to the CUL2-E3 ligase complex for ubiquitination and proteasomal degradation. By using synthetic monobodies that selectively bind the protein tyrosine phosphatase SHP2 and a camelid-derived VHH nanobody that selectively binds the human ASC protein, we demonstrate highly efficient AdPROM-mediated degradation of endogenous SHP2 and ASC in human cell lines. We show that AdPROM-mediated loss of SHP2 in cells impacts SHP2 biology. This study demonstrates for the first time that small polypeptide binders that selectively recognize endogenous target proteins can be exploited for AdPROM-mediated destruction of the target proteins.

The effects of prophylactic iron given in prenatal supplements on iron status and birth outcomes: a randomized controlled trial.

OBJECTIVE: The hypothesis that daily use of a prenatal supplement with iron from enrollment to third trimester to initially iron-replete, nonanemic pregnant women would reduce third-trimester anemia and improve birth outcomes was tested. STUDY DESIGN: Eight hundred sixty-seven women in Raleigh, North Carolina, who were at < 20 weeks of gestation were enrolled; 429 of these women had hemoglobin levels of > or = 110 g/L and ferritin levels of > or = 40 microg/L and were assigned randomly to receive prenatal supplements with 30 mg of iron as ferrous sulfate (n = 218 women) or 0 mg of iron (n = 211 women) until 26 to 29 weeks of gestation. Intent-to-treat analysis was used for the outcomes of third-trimester iron status, birth weight, preterm birth, and small-for-gestational age. RESULTS: Mean birth weight was higher by 108 g (P = .03), and the incidence of preterm delivery was lower (8% vs 14%; P = .05) in the 30-mg group compared with the control group, respectively. Iron supplementation did not affect the prevalence of small-for-gestational age infants or third-trimester iron status. CONCLUSION: Prophylactic iron supplementation that is begun early in pregnancy among low income women in the United States may have benefits beyond the reduction of iron deficiency anemia during pregnancy.