Lipid nanoparticle-mediated messenger RNA delivery for ex vivo engineering of natural killer cells

Natural killer (NK) cells participate in the immune system by eliminating cancer and virally infected cells through germline-encoded surface receptors. Their independence from prior activation as well as their significantly lower toxicity have placed them in the spotlight as an alternative to T cells for adoptive cell therapy (ACT). Engineering NK cells with mRNA has shown great potential in ACT by enhancing their tumor targeting and cytotoxicity. However, mRNA transfection of NK cells is challenging, as the most common delivery methods, such as electroporation, show limitations. Therefore, an alternative non-viral delivery system that enables high mRNA transfection efficiency with preservation of the cell viability would be beneficial for the development of NK cell therapies. In this study, we investigated both polymeric and lipid nanoparticle (LNP) formulations for eGFP-mRNA delivery to NK cells, based on a dimethylethanolamine and diethylethanolamine polymeric library and on different ionizable lipids, respectively. The mRNA nanoparticles based on cationic polymers showed limited internalization by NK cells and low transfection efficiency. On the other hand, mRNA-LNP formulations were optimized by tailoring the lipid composition and the microfluidic parameters, resulting in a high transfection efficiency (∼100%) and high protein expression in NK cells. In conclusion, compared to polyplexes and electroporation, the optimized LNPs show a greater transfection efficiency and higher overall eGFP expression, when tested in NK (KHYG-1) and T (Jurkat) cell lines, and cord blood-derived NK cells. Thus, LNP-based mRNA delivery represents a promising strategy to further develop novel NK cell therapies

Elliptic flow of electrons from heavy-flavor hadron decays in Au+Au collisions at sNN=\sqrt{s_{\rm NN}} = 200, 62.4, and 39 GeV

We present measurements of elliptic flow ($v_2$) of electrons from the decays of heavy-flavor hadrons ($e_{HF}$) by the STAR experiment. For Au+Au collisions at $\sqrt{s_{\rm NN}} =$ 200 GeV we report $v_2$, for transverse momentum ($p_T$) between 0.2 and 7 GeV/c using three methods: the event plane method ($v_{2}${EP}), two-particle correlations ($v_2${2}), and four-particle correlations ($v_2${4}). For Au+Au collisions at $\sqrt{s_{\rm NN}}$ = 62.4 and 39 GeV we report $v_2${2} for $p_T< 2$ GeV/c. $v_2${2} and $v_2${4} are non-zero at low and intermediate $p_T$ at 200 GeV, and $v_2${2} is consistent with zero at low $p_T$ at other energies. The $v_2${2} at the two lower beam energies is systematically lower than at $\sqrt{s_{\rm NN}} =$ 200 GeV for $p_T < 1$ GeV/c. This difference may suggest that charm quarks interact less strongly with the surrounding nuclear matter at those two lower energies compared to $\sqrt{s_{\rm NN}} = 200$ GeV.Comment: Version accepted by PR

Centrality dependence of identified particle elliptic flow in relativistic heavy ion collisions at sqrt(s)= 7.7--62.4 GeV

Elliptic flow (v_2) values for identified particles at midrapidity in Au + Au collisions measured by the STAR experiment in the Beam Energy Scan at the Relativistic Heavy Ion Collider at sqrt{s_{NN}}= 7.7--62.4 GeV are presented for three centrality classes. The centrality dependence and the data at sqrt{s_{NN}}= 14.5 GeV are new. Except at the lowest beam energies we observe a similar relative v_2 baryon-meson splitting for all centrality classes which is in agreement within 15% with the number-of-constituent quark scaling. The larger v_2 for most particles relative to antiparticles, already observed for minimum bias collisions, shows a clear centrality dependence, with the largest difference for the most central collisions. Also, the results are compared with A Multiphase Transport Model and fit with a Blast Wave model.Comment: 14 pages, 12 figures, Phys. Rev. C, to be published. Data tables available at https://drupal.star.bnl.gov/STAR/publications/centrality-dependence-identified-particle-elliptic-flow-relativistic-heavy-ion-collisi

Beam Energy Dependence of the Third Harmonic of Azimuthal Correlations in Au+Au Collisions at RHIC

We present results from a harmonic decomposition of two-particle azimuthal correlations measured with the STAR detector in Au+Au collisions for energies ranging from $\sqrt{s_{NN}}=7.7$ GeV to 200 GeV. The third harmonic $v_3^2\{2\}=\langle \cos3(\phi_1-\phi_2)\rangle$, where $\phi_1-\phi_2$ is the angular difference in azimuth, is studied as a function of the pseudorapidity difference between particle pairs $\Delta\eta = \eta_1-\eta_2$. Non-zero {\vthree} is directly related to the previously observed large-$\Delta\eta$ narrow-$\Delta\phi$ ridge correlations and has been shown in models to be sensitive to the existence of a low viscosity Quark Gluon Plasma (QGP) phase. For sufficiently central collisions, $v_3^2\{2\}$ persist down to an energy of 7.7 GeV suggesting that QGP may be created even in these low energy collisions. In peripheral collisions at these low energies however, $v_3^2\{2\}$ is consistent with zero. When scaled by pseudorapidity density of charged particle multiplicity per participating nucleon pair, $v_3^2\{2\}$ for central collisions shows a minimum near {\snn}$=20$ GeV.Comment: 7 pages, 4 figures, for submission to Phys. Rev. Let