Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors

While induced pluripotent stem cells (iPSCs) hold great clinical promise, one hurdle that remains is the existence of a parental germ-layer memory in reprogrammed cells leading to preferential differentiation fates. While it is problematic for generating cells vastly different from the reprogrammed cells\u27 origins, it could be advantageous for the reliable generation of germ-layer specific cell types for future therapeutic use. Here we use human osteoblast-derived iPSCs (hOB-iPSCs) to generate induced osteoprogenitors (iOPs). Osteoblasts were successfully reprogrammed and demonstrated by endogenous upregulation of Oct4, Sox2, Nanog, TRA-1-81, TRA-16-1, SSEA3, and confirmatory hPSC Scorecard Algorithmic Assessment. The hOB-iPSCs formed embryoid bodies with cells of ectoderm and mesoderm but have low capacity to form endodermal cells. Differentiation into osteoprogenitors occurred within only 2-6 days, with a population doubling rate of less than 24 hrs; however, hOB-iPSC derived osteoprogenitors were only able to form osteogenic and chondrogenic cells but not adipogenic cells. Consistent with this, hOB-iOPs were found to have higher methylation of PPAR gamma but similar levels of methylation on the RUNX2 promoter. These data demonstrate that iPSCs can be generated from human osteoblasts, but variant methylation patterns affect their differentiation capacities. Therefore, epigenetic memory can be exploited for efficient generation of clinically relevant quantities of osteoprogenitor cells

Extreme Telomere Length Dimorphism in the Tasmanian Devil and Related Marsupials Suggests Parental Control of Telomere Length

Telomeres, specialised structures that protect chromosome ends, play a critical role in preserving chromosome integrity. Telomere dynamics in the Tasmanian devil (Sarcophilus harrisii) are of particular interest in light of the emergence of devil facial tumour disease (DFTD), a transmissible malignancy that causes rapid mortality and threatens the species with extinction. We used fluorescent in situ hybridisation to investigate telomere length in DFTD cells, in healthy Tasmanian devils and in four closely related marsupial species. Here we report that animals in the Order Dasyuromorphia have chromosomes characterised by striking telomere length dimorphism between homologues. Findings in sex chromosomes suggest that telomere length dimorphism may be regulated by events in the parental germlines. Long telomeres on the Y chromosome imply that telomere lengthening occurs during spermatogenesis, whereas telomere diminution occurs during oogenesis. Although found in several somatic cell tissue types, telomere length dimorphism was not found in DFTD cancer cells, which are characterised by uniformly short telomeres. This is, to our knowledge, the first report of naturally occurring telomere length dimorphism in any species and suggests a novel strategy of telomere length control. Comparative studies in five distantly related marsupials and a monotreme indicate that telomere dimorphism evolved at least 50 million years ago. © 2012 Bender et al

Aperture, A Large Telescope Using Magnetostriction For Post Deployment Corrections: Final Technical Report of NASA Innovative Advanced Concepts (NIAC) Phase I

In summary, astronomical as well as Earth observing applications of the future are counting on larger aperture telescopes than are currently available. Several groups have been working on the topic of enabling large (about 16-m diameter) UV-Vis telescopes for many years. The unique feature of our concept is that magnetic films are used rather than electrostatic films or piezo-electrostatic pads. Our magnetic film concept allows for contiguous correction along the surface, does not require a hard wire connection, and does not require continuous external application of the field. There are many unknowns related to the initial accuracy of the deployed figure prior to the magnetic write head corrections. The length scale over which the corrections need to be applied is also of concern. For, although approximately mm length scale corrections can be made with the MSM plus write head technology, the number of 1 mm patches in a 16 m diameter mirror is too large to contemplate applying individual corrections to each individual patch. However, deployment strategies and the materials available continue to evolve, in particular shape memory composites (SMCs) [34] or alloys (SMAs) [41], such that at this time we see no show-stoppers for this concept. Furthermore, the ability to tune deformations down to much (factors of 10-100) smaller (m) scale opens the futuristic possibility of improving the figure well beyond Strehl values of 90%

Implementation of a Central Sensorimotor Integration Test for Characterization of Human Balance Control During Stance

Balance during stance is regulated by active control mechanisms that continuously estimate body motion, via a “sensory integration” mechanism, and generate corrective actions, via a “sensory-to-motor transformation” mechanism. The balance control system can be modeled as a closed-loop feedback control system for which appropriate system identification methods are available to separately quantify the sensory integration and sensory-to-motor components of the system. A detailed, functionally meaningful characterization of balance control mechanisms has potential to improve clinical assessment and to provide useful tools for answering clinical research questions. However, many researchers and clinicians do not have the background to develop systems and methods appropriate for performing identification of balance control mechanisms. The purpose of this report is to provide detailed information on how to perform what we refer to as “central sensorimotor integration” (CSMI) tests on a commercially available balance test device (SMART EquiTest CRS, Natus Medical Inc, Seattle WA) and then to appropriately analyze and interpret results obtained from these tests. We describe methods to (1) generate pseudorandom stimuli that apply cyclically-repeated rotations of the stance surface and/or visual surround (2) measure and calibrate center-of-mass (CoM) body sway, (3) calculate frequency response functions (FRFs) that quantify the dynamic characteristics of stimulus-evoked CoM sway, (4) estimate balance control parameters that quantify sensory integration by measuring the relative contribution of different sensory systems to balance control (i.e., sensory weights), and (5) estimate balance control parameters that quantify sensory-to-motor transformation properties (i.e., feedback time delay and neural controller stiffness and damping parameters). Additionally, we present CSMI test results from 40 subjects (age range 21–59 years) with normal sensory function, 2 subjects with results illustrating deviations from normal balance function, and we summarize results from previous studies in subjects with vestibular deficits. A bootstrap analysis was used to characterize confidence limits on parameters from CSMI tests and to determine how test duration affected the confidence with which parameters can be measured. Finally, example results are presented that illustrate how various sensory and central balance deficits are revealed by CSMI testing

VGLL2-NCOA2 leverages developmental programs for pediatric sarcomagenesis

Clinical sequencing efforts are rapidly identifying sarcoma gene fusions that lack functional validation. An example is the fusion of transcriptional coactivators, VGLL2-NCOA2, found in infantile rhabdomyosarcoma. To delineate VGLL2-NCOA2 tumorigenic mechanisms and identify therapeutic vulnerabilities, we implement a cross-species comparative oncology approach with zebrafish, mouse allograft, and patient samples. We find that VGLL2-NCOA2 is sufficient to generate mesenchymal tumors that display features of immature skeletal muscle and recapitulate the human disease. A subset of VGLL2-NCOA2 zebrafish tumors transcriptionally cluster with embryonic somitogenesis and identify VGLL2-NCOA2 developmental programs, including a RAS family GTPase, ARF6. In VGLL2-NCOA2 zebrafish, mouse, and patient tumors, ARF6 is highly expressed. ARF6 knockout suppresses VGLL2-NCOA2 oncogenic activity in cell culture, and, more broadly, ARF6 is overexpressed in adult and pediatric sarcomas. Our data indicate that VGLL2-NCOA2 is an oncogene that leverages developmental programs for tumorigenesis and that reactivation or persistence of ARF6 could represent a therapeutic opportunity