Cell-penetrating peptides as transporters for morpholino oligomers: effects of amino acid composition on intracellular delivery and cytotoxicity

Arginine-rich cell-penetrating peptides (CPPs) are promising transporters for intracellular delivery of antisense morpholino oligomers (PMO). Here, we determined the effect of L-arginine, D-arginine and non-α amino acids on cellular uptake, splice-correction activity, cellular toxicity and serum binding for 24 CPP−PMOs. Insertion of 6-aminohexanoic acid (X) or β-alanine (B) residues into oligoarginine R8 decreased the cellular uptake but increased the splice-correction activity of the resulting compound, with a greater increase for the sequences containing more X residues. Cellular toxicity was not observed for any of the conjugates up to 10 μM. Up to 60 μM, only the conjugates with ⩾ 5 Xs exhibited time- and concentration-dependent toxicity. Substitution of L-arginine with D-arginine did not increase uptake or splice-correction activity. High concentration of serum significantly decreased the uptake and splice-correction activity of oligoarginine conjugates, but had much less effect on the conjugates containing X or B. In summary, incorporation of X/B into oligoarginine enhanced the antisense activity and serum-binding profile of CPP−PMO. Toxicity of X/B-containing conjugates was affected by the number of Xs, treatment time and concentration. More active, stable and less toxic CPPs can be designed by optimizing the position and number of R, D-R, X and B residues

Astro2020 APC White Paper: The Early Career Perspective on the Coming Decade, Astrophysics Career Paths, and the Decadal Survey Process

In response to the need for the Astro2020 Decadal Survey to explicitly engage early career astronomers, the National Academies of Sciences, Engineering, and Medicine hosted the Early Career Astronomer and Astrophysicist Focus Session (ECFS) on October 8-9, 2018 under the auspices of Committee of Astronomy and Astrophysics. The meeting was attended by fifty six pre-tenure faculty, research scientists, postdoctoral scholars, and senior graduate students, as well as eight former decadal survey committee members, who acted as facilitators. The event was designed to educate early career astronomers about the decadal survey process, to solicit their feedback on the role that early career astronomers should play in Astro2020, and to provide a forum for the discussion of a wide range of topics regarding the astrophysics career path. This white paper presents highlights and themes that emerged during two days of discussion. In Section 1, we discuss concerns that emerged regarding the coming decade and the astrophysics career path, as well as specific recommendations from participants regarding how to address them. We have organized these concerns and suggestions into five broad themes. These include (sequentially): (1) adequately training astronomers in the statistical and computational techniques necessary in an era of "big data", (2) responses to the growth of collaborations and telescopes, (3) concerns about the adequacy of graduate and postdoctoral training, (4) the need for improvements in equity and inclusion in astronomy, and (5) smoothing and facilitating transitions between early career stages. Section 2 is focused on ideas regarding the decadal survey itself, including: incorporating early career voices, ensuring diverse input from a variety of stakeholders, and successfully and broadly disseminating the results of the survey

The early career perspective on the coming decade, astrophysics career paths, and the decadal survey process

We discuss the concerns and ideas that emerged during discussion at the Early Career Focus Session at the National Academies regarding (1) the coming decade and the astrophysics career path, and (2) the decadal survey implementation and dissemination.Publisher PDFPeer reviewe

Functional and Phenotypic Characterization of Siglec-6 on Human Mast Cells

Mast cells are tissue-resident cells that contribute to allergic diseases, among others, due to excessive or inappropriate cellular activation and degranulation. Therapeutic approaches to modulate mast cell activation are urgently needed. Siglec-6 is an immunoreceptor tyrosine-based inhibitory motif (ITIM)-bearing receptor selectively expressed by mast cells, making it a promising target for therapeutic intervention. However, the effects of its engagement on mast cells are poorly defined. Siglec-6 expression and endocytosis on primary human mast cells and mast cell lines were assessed by flow cytometry. SIGLEC6 mRNA expression was examined by single-cell RNAseq in esophageal tissue biopsy samples. The ability of Siglec-6 engagement or co-engagement to prevent primary mast cell activation was determined based on assessments of mediator and cytokine secretion and degranulation markers. Siglec-6 was highly expressed by all mast cells examined, and the SIGLEC6 transcript was restricted to mast cells in esophageal biopsy samples. Siglec-6 endocytosis occurred with delayed kinetics relative to the related receptor Siglec-8. Co-crosslinking of Siglec-6 with FcεRIα enhanced the inhibition of mast cell activation and diminished downstream ERK1/2 and p38 phosphorylation. The selective, stable expression and potent inhibitory capacity of Siglec-6 on human mast cells are favorable for its use as a therapeutic target in mast cell-driven diseases