Radiation gastrointestinal (GI) syndrome is a major lethal toxicity that may occur after a radiation/nuclear incident. Currently, there are no prophylactic countermeasures against radiation GI syndrome lethality for first responders, military personnel, or remediation workers entering a contaminated area. The pathophysiology of this syndrome requires depletion of stem cell clonogens (SCCs) within the crypts of Lieberkühn, which are a subset of cells necessary for postinjury regeneration of gut epithelium. Recent evidence indicates that SCC depletion is not exclusively a result of DNA damage but is critically coupled to ceramide-induced endothelial cell apoptosis within the mucosal microvascular network. Here we show that ceramide generated on the surface of endothelium coalesces to form ceramide-rich platforms that transmit an apoptotic signal. Moreover, we report the generation of 2A2, an anti-ceramide monoclonal antibody that binds to ceramide to prevent platform formation on the surface of irradiated endothelial cells of the murine GI tract. Consequently, we found that 2A2 protected against endothelial apoptosis in the small intestinal lamina propria and facilitated recovery of crypt SCCs, preventing the death of mice from radiation GI syndrome after high radiation doses. As such, we suggest that 2A2 represents a prototype of a new class of anti-ceramide therapeutics and an effective countermeasure against radiation GI syndrome mortality.

The limited localization and penetration of monoclonal antibodies (mAb) into solid tumors restricts their antitumor efficacy. Here, we describe a solid tumor-targeting antibody with enhanced tumor penetration activity. We designed a 22-residue peptide (A22p), which was extracted from the C-terminal basic region of semaphorin 3A (Sema3A) but modified to have higher affinity with neuropilin receptors (NRP), and genetically fused it to the C-terminus of Fc of human immunoglobulin G1 via a 15-residue (G 4 S) 3 linker, generating FcA22p, for the bivalent binding to NRPs. In contrast to Fc or the monovalent A22p peptide alone, Fc-A22p homed to tumor vessels and induced vascular permeability through VE-cadherin downregulation and penetrated tumor tissues by interacting with NRPs in mice bearing human tumor xenografts. We extended the Fc-A22p platform by generating mAb-A22p antibodies of two clinically approved solid tumor-targeting mAbs, the anti-EGF receptor mAb cetuximab (erbitux), and the anti-Her2 mAb trastuzumab (herceptin). The mAb-A22p antibodies retained the intrinsic antigen binding, natural Fc-like biophysical properties, and productivity in mammalian cell cultures, comparable with those of the parent mAbs. In mouse xenograft tumor models, the mAb-A22p antibodies more efficiently homed to tumor vessels and spread into the extravascular tumor parenchyma, which significantly enhanced antitumor efficacy compared with the parent mAbs. Our results suggest that mAb-A22p is a superior format for solid tumor-targeting antibodies due to its enhanced tumor tissue penetration and greater antitumor efficacy compared with conventional mAbs.

The nucleation behavior of Ru deposited by atomic layer deposition (ALD) using bis(ethylcyclopentadienyl)ruthenium precursor and O2 reactant is investigated as a function of the number of ALD cycles. The substrates are thermally grown SiO2, NH3 plasma-treated SiO2, and chemical vapor deposited SiNx. The nucleation of Ru strongly depends on the substrate and is much enhanced on the nitride substrates. Transmission electron microscopy analysis reveals that the maximum density of the nuclei is 5.7×1010cm−2 on the SiO2 surface at 500 ALD cycles, 1.2×1012cm−2 on SiNx at 160 ALD cycles, and 2.3×1012cm−2 on NH3 plasma-nitrided SiO2 at 110 ALD cycles. Although the kinetics of Ru nucleation is different on the various substrates, the overall nucleation process in each case consists of an initial slow nucleation stage and a subsequent fast nucleation stage before the coalescence of the nuclei occurs. Considering the adsorption of Ru precursor on the substrate and the surface diffusion of deposited Ru during an ALD cycle, we suggest a model for describing the nucleation of an ALD film at the initial stage with a low surface coverage based on the atomistic nucleation theory of a thin film. The proposed model shows that the density of the nuclei is proportional to the (i+2)th power of the number of ALD cycles and (i+1)th power of the density of atoms deposited per ALD cycle, where i is the critical nuclei size. By applying the proposed model to the experimental results, the critical nuclei size i is found to be 1. The amounts of Ru atoms deposited per ALD cycle on the NH3 plasma-nitrided SiO2 and SiNx are 70 and 24 times larger, respectively, than that on the SiO2 surface. This model quantitatively describes the nucleation kinetics in the ALD system and is verified by a comparison with the experimental results of Ru on various substrates.