Modeling plasticity and dysplasia of pancreatic ductal organoids derived from human pluripotent stem cells

Personalized in vitro models for dysplasia and carcinogenesis in the pancreas have been constrained by insufficient differentiation of human pluripotent stem cells (hPSCs) into the exocrine pancreatic lineage. Here, we differentiate hPSCs into pancreatic duct-like organoids (PDLOs) with morphological, transcriptional, proteomic, and functional characteristics of human pancreatic ducts, further maturing upon transplantation into mice. PDLOs are generated from hPSCs inducibly expressing oncogenic GNAS, KRAS, or KRAS with genetic covariance of lost CDKN2A and from induced hPSCs derived from a McCune-Albright patient. Each oncogene causes a specific growth, structural, and molecular phenotype in vitro. While transplanted PDLOs with oncogenic KRAS alone form heterogenous dysplastic lesions or cancer, KRAS with CDKN2A loss develop dedifferentiated pancreatic ductal adenocarcinomas. In contrast, transplanted PDLOs with mutant GNAS lead to intraductal papillary mucinous neoplasia-like structures. Conclusively, PDLOs enable in vitro and in vivo studies of pancreatic plasticity, dysplasia, and cancer formation from a genetically defined background

Assessment of the PROSPECT safety systems including socio-economic evaluation

This report provides a new methodology for safety benefit assessment of real-world benefit of the Advanced Driver Assistance Systems (ADAS) in terms of saved lives and prevented injuries as well as the resulting monetary benefit for society. This methodology is demonstrated and applied to PROSPECT systems that address potential crashes of passenger cars with vulnerable road users (VRUs) such as pedestrians and cyclists

Mutations and variants of ONECUT1 in diabetes

Genes involved in distinct diabetes types suggest shared disease mechanisms. Here we show that One Cut Homeobox 1 (ONECUT1) mutations cause monogenic recessive syndromic diabetes in two unrelated patients, characterized by intrauterine growth retardation, pancreas hypoplasia and gallbladder agenesis/hypoplasia, and early-onset diabetes in heterozygous relatives. Heterozygous carriers of rare coding variants of ONECUT1 define a distinctive subgroup of diabetic patients with early-onset, nonautoimmune diabetes, who respond well to diabetes treatment. In addition, common regulatory ONECUT1 variants are associated with multifactorial type 2 diabetes. Directed differentiation of human pluripotent stem cells revealed that loss of ONECUT1 impairs pancreatic progenitor formation and a subsequent endocrine program. Loss of ONECUT1 altered transcription factor binding and enhancer activity and NKX2.2/NKX6.1 expression in pancreatic progenitor cells. Collectively, we demonstrate that ONECUT1 controls a transcriptional and epigenetic machinery regulating endocrine development, involved in a spectrum of diabetes, encompassing monogenic (recessive and dominant) as well as multifactorial inheritance. Our findings highlight the broad contribution of ONECUT1 in diabetes pathogenesis, marking an important step toward precision diabetes medicine

A Comprehensive Evaluation Approach for Highly Automated Driving

Since the last decade, development efforts by academia and industry for automated driving functions have increased significantly. Also, the European research project AdaptIVe is looking into this topic. Due to the large operation spaces and various complex situations that are covered by these functions, efforts for evaluation increase also significantly. Within AdaptIVe, a comprehensive evaluation approach for automated driving functions ranging from SAE level 2-4 has been developed [1]. The approach splits the evaluation into technical, user-related, in-traffic and impact assessment addressing safety and environmental effects of automated driving. For each evaluation type appropriate test tools and methods are selected e.g. field test for technical assessment, trials on test track and in real traffic for the user-related assessments and simulations for the in-traffic and impact assessment. Next to the assessment type also the characteristics of the function must be considered when deciding for specific test tools. Hence, besides to the level of automation [8] the automated driving functions are classified into continuous and event-based operating functions. Whereas event-based operating functions are only operating for a short period in time (e.g. automated parking), continuous operating functions are, once they are active, operating for longer time periods (e.g. highway automation). Based on the classification the aspects to be evaluated and test methods are selected for all assessment types. The developed methodology has been applied to several automated driving functions developed within AdaptIVe. As an example, for the technical assessment of continuous operating functions it has been assessed whether the driving behavior of the developed functions is similar to human driving behavior and therefore not disturbing human traffic. In the user-related assessment, issues related to driver behavior, understanding of automation, trust, mental workload, resuming control, vigilance, usability and acceptance has been looked at. In this paper the key aspects of the AdaptIVe evaluation methodology for technical, user-related, in-traffic and impact assessment are presented as well as the key results of the application of this methodology on the within AdaptIVe developed automated driving functions

A Comprehensive Evaluation Approach for Highly Automated Driving

Since the last decade, development efforts by academia and industry for automated driving functions have increased significantly. Also, the European research project AdaptIVe is looking into this topic. Due to the large operation spaces and various complex situations that are covered by these functions, efforts for evaluation increase also significantly. Within AdaptIVe, a comprehensive evaluation approach for automated driving functions ranging from SAE level 2-4 has been developed [1]. The approach splits the evaluation into technical, user-related, in-traffic and impact assessment addressing safety and environmental effects of automated driving. For each evaluation type appropriate test tools and methods are selected e.g. field test for technical assessment, trials on test track and in real traffic for the user-related assessments and simulations for the in-traffic and impact assessment. Next to the assessment type also the characteristics of the function must be considered when deciding for specific test tools. Hence, besides to the level of automation [8] the automated driving functions are classified into continuous and event-based operating functions. Whereas event-based operating functions are only operating for a short period in time (e.g. automated parking), continuous operating functions are, once they are active, operating for longer time periods (e.g. highway automation). Based on the classification the aspects to be evaluated and test methods are selected for all assessment types. The developed methodology has been applied to several automated driving functions developed within AdaptIVe. As an example, for the technical assessment of continuous operating functions it has been assessed whether the driving behavior of the developed functions is similar to human driving behavior and therefore not disturbing human traffic. In the user-related assessment, issues related to driver behavior, understanding of automation, trust, mental workload, resuming control, vigilance, usability and acceptance has been looked at. In this paper the key aspects of the AdaptIVe evaluation methodology for technical, user-related, in-traffic and impact assessment are presented as well as the key results of the application of this methodology on the within AdaptIVe developed automated driving functions

Deliverable D.6.2 Safety and Economic Benefits - Project PIONEERS - Protective Innovations of New Equipment for Enhanced Rider Safety

The main goal of the PIONEERS project is to improve the safety of Powered-Two-Wheelers by providing an integrated approach to rider protection considering on-rider (Personal Protective Equipment) and on-board systems. The implementation of the PIONEERS' main results will contribute to reducing PTW fatalities and injuries by defining test methods to develop protective systems and on-board systems to reduce impact severity. In order to assess if this major conclusion of the PIONEERS project is being fulfilled and to quantify the benefit, an Impact Evaluation has been developed in this document. This evaluation concerns both economical and safety benefits (in terms of avoided or mitigated accidents, reduction of morbidity and severity of injuries) of the following proposed PTW safety countermeasures that have been developed in PIONEERS: PreCrash Braking System, Airbag jacket and the PTW-PPE communication system, Motorcycle and scooter leg protector. Concerning the PCB, the evaluation was obtained via computer simulations of a set of 60 real-world in-depth crashes. The effects were assessed in terms of a reduction of the absolute and relative impact speed of the PTW. A parametric approach in which PCB intervention parameters were varied (field of view, range, deceleration, fade-in jerk, triggering strategy...) was adopted to compute the effects of the system for different conditions. Such approach led to the identification of three combinations of parameters to represent typical system effects assuming a pessimistic (low efficiency), average, and an optimistic approach (high efficiency). Depending on the set of parameters, benefits in terms of speed reduction can go until a median value of 15km/h. Then societal benefits have been calculated in terms of casualties' reduction or crashes using Injury Risk Function and the new relative impact speed distribution. Results show that global benefits for slight, serious or fatal injuries are included between -4% to -31%. Finally, economic benefits in terms of cost were evaluated using the SafetyCube software. Several configurations were considered like the implementation rate of PCB among all motorcycles in Europe, the cost to promote such systems, the horizon, etc. In a 5 years period, this cost will be balanced by savings from health and social expenses, and in a high rate of implementation (with the average evaluation) Europe could save, at least, 30 human lives in this period. With an optimistic configuration, the net value of the benefits could reached between approximately 500k€ until more than 90 M€. Concerning the Airbag jackets, the societal benefits calculation was first based on the establishment of Injury Risk Function for slight, serious and fatal injuries on the trunk without the airbag. In order to establish these IRF, four accident databases were considered and 382 accidents concerned a rider with at least one injury in trunk body region. Then, according to the work performed in others WP's of the Pioneers project, three hypotheses have been considered for the level of protection and the reduction of injuries. They considered that the airbag jacket provides a reduction of an AIS-1 for speed impact lower than 20km/h, respectively 30km/h and 40km/h. Results of the societal evaluation show that between 1,3% until 19% of injuries could be avoided in function of the configuration. From an economical point of view, with a high implementation rate (6%) in the next 5 years, a net value of more than 140M€ could be saved. Concerning the Lateral Protectors, Injury Risk Function of the Lower Leg in lateral impact configuration were first established based on 81 accidents. Then, based on the findings from the lateral protection devices that have been developed in the PIONEERS project (in particular WP3), two hypotheses have been retained for the level of protection of such systems: one considered as a « low-speed » countermeasure (only effective until 15km/h) and one considered as « medium-speed » countermeasure (only effective between 15 to 30km/h). Global societal and economic benefits have been observed but results have to be taken very cautiously. Indeed, the economic evaluation show that an amount of more than 5M€ for the net benefits could be saved with the best configuration. But it is important to remind that these systems are not still available in the market, so these evaluations have to be considered as a theoretical exercise. In conclusion, this work allowed to evaluate safety and economic benefits of several protective systems: Pre-Crash Braking, airbag jackets, Lateral protectors. But because some of these systems are prototypes and not yet on the market, all the evaluations have to be taken very carefully. In particular, results from societal and economical evaluations are sensible and have to be considered cautiously. Beyond these specific evaluations, theroretical methodologies have been defined and could be applied on other systems

Livrable D1.1 - Powered Two-Wheelers - Road Traffic Accident Scenarios and Common Injuries - Project PIONEERS - Protective Innovations of New Equipment for Enhanced Rider Safety

Comprehensive literature review has been performed related to analyses of road traffic accidents involving powered two-wheelers (PTW). It became obvious that many projects, in particular European research projects, studies from French and Italian research institutes and from Australia have been working on this topic; however, results can hardly be compared since the definitions of many variables vary, different injury coding are used and all analysed in-depth accident datasets are based on greatly differing inclusion criteria. The PIONEERS project took this burden and established definitions for relevant Accident Scenarios and body regions which may form a new common understanding and will accelerate harmonization processes in this research field. Furthermore, several datasets from Europe and Australia (Compilation of macrostatistical European accident data as well as data from national statistics and in-depth accident investigations) of latest years have been analysed to provide a current understanding of the accident occurrence of powered two-wheelers

Transcriptional changes and the role of ONECUT1 in hPSC pancreatic differentiation

Cell type specification during pancreatic development is tightly controlled by a transcriptional and epigenetic network. The precise role of most transcription factors, however, has been only described in mice. To convey such concepts to human pancreatic development, alternative model systems such as pancreatic in vitro differentiation of human pluripotent stem cells can be employed. Here, we analyzed stage-specific RNA-, ChIP-, and ATAC-sequencing data to dissect transcriptional and regulatory mechanisms during pancreatic development. Transcriptome and open chromatin maps of pancreatic differentiation from human pluripotent stem cells provide a stage-specific pattern of known pancreatic transcription factors and indicate ONECUT1 as a crucial fate regulator in pancreas progenitors. Moreover, our data suggest that ONECUT1 is also involved in preparing pancreatic progenitors for later endocrine specification. The dissection of the transcriptional and regulatory circuitry revealed an important role for ONECUT1 within such network and will serve as resource to study human development and disease

Functional genomic screening in human pluripotent stem cells reveals new roadblocks in early pancreatic endoderm formation

Human pluripotent stem cells, with their ability to proliferate indefinitely and to differentiate into virtually all cell types of the human body, provide a novel resource to study human development and to implement relevant disease models. Here, we employed a human pancreatic differentiation platform complemented with an shRNA screen in human pluripotent stem cells (PSCs) to identify potential drivers of early endoderm and pancreatic development. Deep sequencing followed by abundancy ranking pinpointed six top hit genes potentially associated with either improved or impaired endodermal differentiation, which were selected for functional validation in CRISPR-Cas9 mediated knockout (KO) lines. Upon endoderm differentiation (DE), particularly the loss of SLC22A1 and DSC2 led to impaired differentiation efficiency into CXCR4/KIT-positive DE cells. qPCR analysis also revealed changes in differentiation markers CXCR4, FOXA2, SOX17, and GATA6. Further differentiation of PSCs to the pancreatic progenitor (PP) stage resulted in a decreased proportion of PDX1/NKX6-1-positive cells in SLC22A1 KO lines, and in DSC2 KO lines when differentiated under specific culture conditions. Taken together, our study reveals novel genes with potential roles in early endodermal development