Evaluation of TH-Cre knock-in cell lines for detection and specific targeting of stem cell-derived dopaminergic neurons

The focal and progressive degeneration of dopaminergic (DA) neurons in ventral midbrain has made Parkinson's disease (PD) a particularly interesting target of cell-based therapies. However, ethical issues and limited tissue availability have so far hindered the widespread use of human fetal tissue in cell-replacement therapy. DA neurons derived from human pluripotent stem cells (hPSCs) offer unprecedented opportunities to access a renewable source of cells suitable for PD therapeutic applications. To better understand the development and functional properties of stem-cell derived DA neurons, we generated targeted hPSC lines with the gene coding for Cre recombinase knocked into the TH locus. When combined with flexed GFP, they serve as reporter cell lines able to identify and isolate TH+ neurons in vitro and after transplantation in vivo. These TH-Cre lines provide a valuable genetic tool to manipulate DA neurons useful for the design of more precise DA differentiation protocols and the study of these cells after transplantation in pre-clinical animal models of PD

Single-Cell Profiling of Coding and Noncoding Genes in Human Dopamine Neuron Differentiation

Dopaminergic (DA) neurons derived from human pluripotent stem cells (hPSCs) represent a renewable and available source of cells useful for understanding development, developing disease models, and stem-cell therapies for Parkinson's disease (PD). To assess the utility of stem cell cultures as an in vitro model system of human DA neurogenesis, we performed high-throughput transcriptional profiling of ~20,000 ventral midbrain (VM)-patterned stem cells at different stages of maturation using droplet-based single-cell RNA sequencing (scRNAseq). Using this dataset, we defined the cellular composition of human VM cultures at different timepoints and found high purity DA progenitor formation at an early stage of differentiation. DA neurons sharing similar molecular identities to those found in authentic DA neurons derived from human fetal VM were the major cell type after two months in culture. We also developed a bioinformatic pipeline that provided a comprehensive long noncoding RNA landscape based on temporal and cell-type specificity, which may contribute to unraveling the intricate regulatory network of coding and noncoding genes in DA neuron differentiation. Our findings serve as a valuable resource to elucidate the molecular steps of development, maturation, and function of human DA neurons, and to identify novel candidate coding and noncoding genes driving specification of progenitors into functionally mature DA neurons

Grafts Derived from an α-Synuclein Triplication Patient Mediate Functional Recovery but Develop Disease-Associated Pathology in the 6-OHDA Model of Parkinson's Disease

BACKGROUND: Human induced pluripotent stem cells (hiPSCs) have been proposed as an alternative source for cell replacement therapy for Parkinson's disease (PD) and they provide the option of using the patient's own cells. A few studies have investigated transplantation of patient-derived dopaminergic (DA) neurons in preclinical models; however, little is known about the long-term integrity and function of grafts derived from patients with PD.OBJECTIVE: To assess the viability and function of DA neuron grafts derived from a patient hiPSC line with an α-synuclein gene triplication (AST18), using a clinical grade human embryonic stem cell (hESC) line (RC17) as a reference control.METHODS: Cells were differentiated into ventral mesencephalic (VM)-patterned DA progenitors using an established GMP protocol. The progenitors were then either terminally differentiated to mature DA neurons in vitro or transplanted into 6-hydroxydopamine (6-OHDA) lesioned rats and their survival, maturation, function, and propensity to develop α-synuclein related pathology, were assessed in vivo.RESULTS: Both cell lines generated functional neurons with DA properties in vitro. AST18-derived VM progenitor cells survived transplantation and matured into neuron-rich grafts similar to the RC17 cells. After 24 weeks, both cell lines produced DA-rich grafts that mediated full functional recovery; however, pathological changes were only observed in grafts derived from the α-synuclein triplication patient line.CONCLUSION: This data shows proof-of-principle for survival and functional recovery with familial PD patient-derived cells in the 6-OHDA model of PD. However, signs of slowly developing pathology warrants further investigation before use of autologous grafts in patients

The Kepler-10 planetary system revisited by HARPS-N: A hot rocky world and a solid Neptune-mass planet

Kepler-10b was the first rocky planet detected by the Kepler satellite and con- firmed with radial velocity follow-up observations from Keck-HIRES. The mass of the planet was measured with a precision of around 30%, which was insufficient to constrain models of its internal structure and composition in detail. In addition to Kepler-10b, a second planet transiting the same star with a period of 45 days was sta- tistically validated, but the radial velocities were only good enough to set an upper limit of 20 Mearth for the mass of Kepler-10c. To improve the precision on the mass for planet b, the HARPS-N Collaboration decided to observe Kepler-10 intensively with the HARPS-N spectrograph on the Telescopio Nazionale Galileo on La Palma. In to- tal, 148 high-quality radial-velocity measurements were obtained over two observing seasons. These new data allow us to improve the precision of the mass determina- tion for Kepler-10b to 15%. With a mass of 3.33 +/- 0.49 Mearth and an updated radius of 1.47 +0.03 -0.02 Rearth, Kepler-10b has a density of 5.8 +/- 0.8 g cm-3, very close to the value -0.02 predicted by models with the same internal structure and composition as the Earth. We were also able to determine a mass for the 45-day period planet Kepler-10c, with an even better precision of 11%. With a mass of 17.2 +/- 1.9 Mearth and radius of 2.35 +0.09 -0.04 Rearth, -0.04 Kepler-10c has a density of 7.1 +/- 1.0 g cm-3. Kepler-10c appears to be the first strong evidence of a class of more massive solid planets with longer orbital periods.Comment: 44 pages, 8 figures, accepted for publication in Ap

Kepler-102 : masses and compositions for a super-Earth and sub-Neptune orbiting an active star

Funding: This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under grant No. 1842402. C.L.B., L.W., and D.H. acknowledge support from National Aeronautics and Space Administration (grant No. 80NSSC19K0597) issued through the Astrophysics Data Analysis Program. D.H. also acknowledges support from the Alfred P. Sloan Foundation. K.R. acknowledges support from the UK STFC via grant No. ST/V000594/1. E.G. acknowledges support from NASA grant No. 80NSSC20K0957 (Exoplanets Research Program).Radial velocity (RV) measurements of transiting multiplanet systems allow us to understand the densities and compositions of planets unlike those in the solar system. Kepler-102, which consists of five tightly packed transiting planets, is a particularly interesting system since it includes a super-Earth (Kepler-102d) and a sub-Neptune-sized planet (Kepler-102e) for which masses can be measured using RVs. Previous work found a high density for Kepler-102d, suggesting a composition similar to that of Mercury, while Kepler-102e was found to have a density typical of sub-Neptune size planets; however, Kepler-102 is an active star, which can interfere with RV mass measurements. To better measure the mass of these two planets, we obtained 111 new RVs using Keck/HIRES and Telescopio Nazionale Galileo/HARPS-N and modeled Kepler-102's activity using quasiperiodic Gaussian process regression. For Kepler-102d, we report a mass upper limit Md < 5.3 M⊕ (95% confidence), a best-fit mass Md = 2.5 ± 1.4 M⊕, and a density ρd = 5.6 ± 3.2 g cm−3, which is consistent with a rocky composition similar in density to the Earth. For Kepler-102e we report a mass Me = 4.7 ± 1.7 M⊕ and a density ρe = 1.8 ± 0.7 g cm−3. These measurements suggest that Kepler-102e has a rocky core with a thick gaseous envelope comprising 2%–4% of the planet mass and 16%–50% of its radius. Our study is yet another demonstration that accounting for stellar activity in stars with clear rotation signals can yield more accurate planet masses, enabling a more realistic interpretation of planet interiors.Publisher PDFPeer reviewe

Independent validation of the temperate super-Earth HD 79211 b using HARPS-N

This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under grant No. DGE1745303. The HARPS-N project was funded by the Prodex Program of the Swiss Space Office (SSO), the Harvard- University Origin of Life Initiative (HUOLI), the Scottish Universities Physics Alliance (SUPA), the University of Geneva, the Smithsonian Astrophysical Observatory (SAO), the Italian National Astrophysical Institute (INAF), University of St. Andrews, Queen's University Belfast, and University of Edinburgh. Parts of this work have been supported by the National Aeronautics and Space Administration under grant No. NNX17AB59G, issued through the Exoplanets Research Program. Parts of this work have been supported by the Brinson Foundation. R.D.H. is funded by the UK Science and Technology Facilities Council (STFC)'s Ernest Rutherford Fellowship (grant No. ST/V004735/1). T.G.W and A.C.C acknowledge support from STFC consolidated grant Nos. ST/R000824/1 and ST/V000861/1, and UKSA grant ST/R003203/1.We present high-precision radial velocities (RVs) from the HARPS-N spectrograph for HD 79210 and HD 79211, two M0V members of a gravitationally bound binary system. We detect a planet candidate with a period of 24.421−0.017+0.016 days around HD 79211 in these HARPS-N RVs, validating the planet candidate originally identified in CARMENES RV data alone. Using HARPS-N, CARMENES, and RVs spanning a total of 25 yr, we further refine the planet candidate parameters to P = 24.422 ± 0.014 days, K = 3.19 ± 0.27 m s−1, M sin i = 10.6 ± 1.2M⊕, and a = 0.142 ± 0.005 au. We do not find any additional planet candidate signals in the data of HD 79211, nor do we find any planet candidate signals in HD 79210. This system adds to the number of exoplanets detected in binaries with M-dwarf members and serves as a case study for planet formation in stellar binaries.Publisher PDFPeer reviewe

Preclinical quality, safety, and efficacy of a human embryonic stem cell-derived product for the treatment of Parkinson’s disease, STEM-PD

Cell replacement therapies for Parkinson’s disease (PD) based on transplantation of pluripotent stem cell-derived dopaminergic neurons are now entering clinical trials. Here, we present quality, safety, and efficacy data supporting the first-in-human STEM-PD phase I/IIa clinical trial along with the trial design. The STEM-PD product was manufactured under GMP and quality tested in vitro and in vivo to meet regulatory requirements. Importantly, no adverse effects were observed upon testing of the product in a 39-week rat GLP safety study for toxicity, tumorigenicity, and biodistribution, and a non-GLP efficacy study confirmed that the transplanted cells mediated full functional recovery in a pre-clinical rat model of PD. We further observed highly comparable efficacy results between two different GMP batches, verifying that the product can be serially manufactured. A fully in vivo-tested batch of STEM-PD is now being used in a clinical trial of 8 patients with moderate PD, initiated in 2022

Kepler-21b: A Rocky Planet Around a V = 8.25 Magnitude Star

HD 179070, aka Kepler-21, is a V = 8.25 F6IV star and the brightest exoplanet host discovered by Kepler. An early detailed analysis by Howell et al. (2012) of the first thirteen months (Q0 - Q5) of Kepler light curves revealed transits of a planetary companion, Kepler-21b, with a radius of about 1.60 ± 0.04 R⊕ and an orbital period of about 2.7857 days. However, they could not determine the mass of the planet from the initial radial velocity observations with Keck-HIRES, and were only able to impose a 2σ upper limit of 10 M⊕. Here we present results from the analysis of 82 new radial velocity observations of this system obtained with HARPS-N, together with the existing 14 HIRES data points. We detect the Doppler signal of Kepler-21b with a radial velocity semi-amplitude K = 2.00 ± 0.65 m s-1, which corresponds to a planetary mass of 5.1 ± 1.7 M⊕. We also measure an improved radius for the planet of 1.639 +0.019/-0.015 R⊕, in agreement with the radius reported by Howell et al. (2012). We conclude that Kepler-21b, with a density of 6.4 ± 2.1 g cm-3, belongs to the population of small, ≤6 M⊕ planets with iron and magnesium silicate interiors, which have lost the majority of their envelope volatiles via stellar winds or gravitational escape. The RV analysis presented in this paper serves as example of the type of analysis that will be necessary to confirm the masses of TESS small planet candidates.PostprintPeer reviewe