CXCR4 peptide-based fluorescence endoscopy in a mouse model of Barrett’s esophagus

Background Near-infrared (NIR) fluorescence imaging has been emerging as a promising strategy to overcome the high number of early esophageal adenocarcinomas missed by white light endoscopy and random biopsy collection. We performed a preclinical assessment of fluorescence imaging and endoscopy using a novel CXCR4-targeted fluorescent peptide ligand in the L2-IL1B mouse model of Barrett’s esophagus. Methods Six L2-IL1B mice with advanced stage of disease (12–16 months old) were injected with the CXCR4-targeted, Sulfo-Cy5-labeled peptide (MK007), and ex vivo wide-field imaging of the whole stomach was performed 4 h after injection. Before ex vivo imaging, fluorescence endoscopy was performed in three L2-IL1B mice (12–14 months old) by a novel imaging system with two L2-IL1B mice used as negative controls. Results Ex vivo imaging and endoscopy in L2-IL1B mice showed that the CXCR4-targeted MK007 accumulated mostly in the dysplastic lesions with a mean target-to-background ratio > 2. The detection of the Sulfo-Cy5 signal in dysplastic lesions and its co-localization with CXCR4 stained cells by confocal microscopy further confirmed the imaging results. Conclusions This preliminary preclinical study shows that CXCR4-targeted fluorescence endoscopy using MK007 can detect dysplastic lesions in a mouse model of Barrett’s esophagus. Further investigations are needed to assess its use in the clinical setting.

Ex-vivo imaging of buccal and oral periodontal bone with low-dose CBCT in porcine jaws

Objectives: New CBCT devices have been developed which can provide “low-dose CBCTs (LD-CBCT)”. Aim of this study is to investigate the suitability of LD-CBCT for measurement of alveolar buccal/oral bone. Methods and materials: Vestibular and oral bone loss of the teeth of seven porcine mandibles free of soft tissues were investigated by Micro-CT and three CBCT-modes: high-dose (HD), standard-dose (SD) and low-dose (LD). Radiographic measurements of bone loss (bl) and vestibular and oral bone thickness (bt) were made by two raters at 69 sites. Measurement means and differences, Intraclass correlation (ICC) and Bland–Altman plots were calculated. Results: ICCs between raters(r) concerning bl were 0.954 for HD, 0.949 for SD and 0.945 for LD; concerning bt they were 0.872 for HD, 0.845 for SD and 0.783 for LD. Means of differences of bt measurements were −0.01 mm(r1)/0.00 mm(r2) for HD, 0.04 mm(r1)/0.02 mm(r2) for SD and 0.02 mm(r1)/0.04 mm(r2) for LD; for bl measurements they were 0.06 mm(r1)/0.05 mm(r2) for HD, −0.01 mm(r1)/0.13 mm(r2) for SD and 0.07 mm(r1)/0.16 mm(r2) for LD. Linear regression indicates no noticeable differences between methods and the raters with respect to bl and bt. Conclusions Relating to the CBCT-device used in this study, LD-CBCT is a promising method to detect and describe buccal and oral periodontal bl and bt. Further studies with human anatomic structures must confirm these results.

Highly Potent GalNAc-Conjugated Tiny LNA Anti-miRNA-122 Antisense Oligonucleotides

The development of clinically relevant anti-microRNA antisense oligonucleotides (anti-miRNA ASOs) remains a major challenge. One promising configuration of anti-miRNA ASOs called “tiny LNA (tiny Locked Nucleic Acid)” is an unusually small (~8-mer), highly chemically modified anti-miRNA ASO with high activity and specificity. Within this platform, we achieved a great enhancement of the in vivo activity of miRNA-122-targeting tiny LNA by developing a series of N-acetylgalactosamine (GalNAc)-conjugated tiny LNAs. Specifically, the median effective dose (ED50) of the most potent construct, tL-5G3, was estimated to be ~12 nmol/kg, which is ~300–500 times more potent than the original unconjugated tiny LNA. Through in vivo/ex vivo imaging studies, we have confirmed that the major advantage of GalNAc over tiny LNAs can be ascribed to the improvement of their originally poor pharmacokinetics. We also showed that the GalNAc ligand should be introduced into its 5′ terminus rather than its 3′ end via a biolabile phosphodiester bond. This result suggests that tiny LNA can unexpectedly be recognized by endogenous nucleases and is required to be digested to liberate the parent tiny LNA at an appropriate time in the body. We believe that our strategy will pave the way for the clinical application of miRNA-targeting small ASO therapy.

Tumor Targeted Nanoparticles Improve Therapeutic Index of BCL2 and MCL1 dual inhibition

Cancer and normal cells utilize multiple anti-apoptotic BCL2 proteins to prevent cell death. Therapeutic targeting of multiple BCL2 family proteins enhances tumor killing, but is also associated with increased systemic toxicity. Here, we demonstrate that the dual targeting of MCL1 and BCL2 proteins, using the small molecules S63845 and venetoclax, induces durable remissions in mice harboring human DLBCL tumors but is accompanied by hematological toxicity and weight loss. To mitigate these toxicities, we encapsulated S63845 or venetoclax into nanoparticles targeting P-selectin that is enriched in tumor endothelial cells. In vivo and ex vivo imaging demonstrated preferential targeting of the nanoparticles to lymphoma tumors over vital organs. Mass-spectrometry analyses after nanoparticle drug administration confirmed tumor enrichment of the drug while reducing plasma levels. Furthermore, nanoparticle encapsulation allowed 3.5 to 6.5-fold reduction in drug dose, induced sustained remissions and minimized toxicity. Our results support the development of nanoparticles to deliver BH3 mimetic combinations in lymphoma and in general for toxic drugs in cancer therapy.