Spaceflight associated neuro-ocular syndrome (SANS) is one of the potential barriers to human long-duration spaceflight (LDSF), including a manned mission to Mars. While a large barrier, the pathophysiology of SANS is not well understood, and functional and structural findings from SANS continue to be further characterized. Currently on the International Space Station (ISS), scheduled visual assessments are static visual acuity, Amsler grid, and a self-reported survey. Additional visual assessments may help the understanding of this neuro-ophthalmic phenomenon, as well as the effects of spaceflight of overall ocular health. In this paper, a case is made for expanding scheduled visual assessments to include dynamic visual, contrast sensitivity (CS), visual field testing, and virtual reality-based metamorphopsia assessment during spaceflight. These further assessments may play a key role in helping to determine the structural and functional changes associated with SANS, which are crucial to maintain astronaut vision during LDSF, as well as for developing countermeasures. Finally, a brief discussion is provided about current challenges to expanding visual testing during spaceflight and potential solutions to these barriers, specifically head-mounted visual assessment technology. The visual and structural impact of SANS occurs in astronauts engaged in longer duration space missions. While a considerable barrier to future spaceflight, the pathophysiology of SANS is poorly defined. In addition, the functional and structural effects of microgravity on the neuro-ophthalmic system continue to be further characterized.Spaceflight associated neuro-ocular syndrome (SANS) is a collection of neuro-ocular imaging and clinical findings, including unilateral and bilateral optic disc edema (ODE), posterior globe flattening, hyperopic refractive error shifts, cotton wool spots, and chorioretinal folds.As these neuro-ocular stresses are not experienced by crews in isolation, it is crucial to consider how they together affect human physiology. The space exposome reflects a person’s total exposure to their surroundings as well as how these exposures interact with other personal characteristics like age, sex, and genetics. It is thus a unifying framework that depicts the interplay of all environmental effects on the human body. This framework will influence how space travel will affect the human system. For these assessments, crewmembers who spend six months in-orbit are evaluated pre-launch, after 30 launch days (LD30), after LD90, and 30 days before returning to Earth. In addition, evaluations may be performed when clinically indicated. For crewmembers who spend one year in-orbit, astronauts on the ISS undergo these assessments at LD30, LD90, LD180, and LD270, in addition to 30 days before returning to Earth. Contrast sensitivity (CS) is available onboard the ISS, but only utilized when clinically indicated. However, scheduled CS assessments during spaceflight may help further characterize SANS and its pathogenesis regarding cephalad fluid shifts and intracranial pressure (ICP).Currently in-orbit onboard the International Space Station (ISS), the only forms of functional visual assessment performed include static visual acuity, Amsler grid, and a self-reported survey. Potentially, ODE can result in enlargement of the blind spot and other visual field scotomas, but there is currently no automated perimeter onboard the ISS. According to the SANS Evidence Report, visual distortions or decreased visual acuity that cannot be treated by glasses or contact lenses is a theoretic risk of SANS. Globe flattening  occurs in around 16% and 29% of crewmembers during short- and long-duration space voyages, respectively, and leads to the refractive hyperopic shift in SANS. Special “Space Anticipation Glasses” with customizable positive lens power settings have been used on the ISS prophylactically to minimize such refractive errors. Currently, however, astronauts bring their own reading add spectacles on the ISS. The clinical findings in astronauts following long-duration spaceflight (LDSF) that include ODE and chorioretinal folds have been investigated using imaging technology such OCT and magnetic resonance imaging (MRI).On the ISS, prior work has demonstrated that 69% of the United States crew members have at least one eye with a > 20µm increase in peripapillary retinal nerve fiber layer thickness (RNFL) on optical coherence tomography (OCT) measurements. This increased RNFL correlates with the ophthalmoscopic sign of ODE. Alongside ODE, other ocular findings in astronauts on long- or short-duration flights (around six months) include refractive changes or “hyperopic shift” and chorioretinal folds.In this paper, a case is made for expanding scheduled visual assessments during spaceflight to include CS, dynamic visual acuity (DVA), visual field perimetry, and head-mounted metamorphopsia assessment. These clinically insightful assessments may play a key role in helping to determine the structural and functional changes associated with SANS, which are crucial to maintain astronaut vision during LDSF, as well as for developing countermeasures. Following 24 hours after return from LDSF, it was previously reported that astronauts had a significantly decreased DVA of 0.75 eye chart lines, with some astronauts performing similarly to individuals with vestibular impairments. As these results occurred 24 hours after landing, the decrease in DVA during and directly following a gravitational (G)-transition event is likely much greater and may pose a significant risk to astronaut mission performance during this critical period. Current research is focused on utilizing head-mounted DVA assessments to evaluate astronaut safety following G-transitions. Further DVA assessments of astronauts in-flight and post-flight are required to further understand these G-transition effects.Dynamic visual acuity is the ability to visualize an object in motion and is essential in a rapidly changing environment such as during spaceflight. In-flight Space Motion Sickness (SMS) has been a major topic of discussion and research at National Aeronautics and Space Administration  recently, since most missions performed are short in duration (around ten days). But LDSF increases the risk of developing re-entry SMS. Onset of in-flight and re-entry motion sickness occurs typically within a few minutes of G-transition, with head movement sensitivity lasting for days. Notably, sudden or triggering head movements, visual reorientation illusions can cause vomiting.Advances in developing this technology is particularly useful as one of the biggest clinical issues affecting crew functions during G-transitions is motion sickness. Testing for CS and other tests are currently only performed on the ISS if clinically indicated. Regular CS testing and formal automated visual field testing should be considered as a component of regular vision testing for all astronauts.Although ODE is one of the most common findings in astronauts after LDSF, the clinical impact of SANS-related ODE on functional visual parameters remains ill defined. The development of ODE on Earth may cause decreased color vision, decreased visual acuity, and constriction of the visual field, but to date, no case of SANS has reported to cause permanent visual acuity or visual field loss. During spaceflight, there is a loss of hydrostatic pressure, leading to cephalad fluid shifts and possible increased ICP. However, several factors in the clinical presentation and post-flight tests have not been consistent with elevated ICP, leading to a name change from VIIP to SANS. However, direct ICP has never been measured during spaceflight due to the invasiveness of a lumbar puncture procedure, the gold standard for ICP measurements. Thus, the role of ICP in SANS is still not well understood. As once termed VIIP, idiopathic intracranial pressure (IIH) has been considered to be a terrestrial analog to SANS. Peripheral CS testing was recently shown to be significantly more affected than central CS in patients with IIH. Evaluating such patterns during spaceflight with CS may give additional insight into the role of ICP in SANS development.Initially termed “Vision Impairment and Intracranial Pressure” (VIIP), SANS was thought to be initially caused by increased ICP.Numerous theories have been put forward to understand why ICP-related changes might happen, including venous stasis and cerebral edema. In addition, the potential role of the recently discovered ocular glymphatic system has also been postulated in SANS. Some authors have hypothesized that cephalad fluid shifts during LDSF can contribute to elevated ICP. The cerebrospinal fluid (CSF) is predominantly produced in the choroid plexus and then drains into the cerebral venous system. Some studies suggest that a spaceflight-induced reduction in CSF drainage into cerebral veins alongside congestion in this system could result in optic nerve sheath expansion and globe flattening. Although SANS shares some clinical and imaging features with terrestrial IIH, many authors have argued that terrestrial IIH is not a good model for SANS. Typically, IIH presents with clinical features that have rarely, if ever, been described among astronauts with SANS .Automated perimetry serves as a highly useful assessment for various conditions that can affect central vision  or peripheral vision . There are established risks for visual field loss after LDSF outside of SANS, including increased risk for atherosclerotic disease during spaceflight, radiation exposure, and cardiovascular effects that could theoretically produce visual loss. Scott, et al concluded in a study on goggles as a countermeasure for SANS that potential compression of the lamina cribrosa may lead to damage of the retinal ganglion cell axons, as observed to terrestrial glaucoma. Thus, if these future countermeasures are employed for future spaceflight, appropriate visual assessments must also be employed to ensure that peripheral vision is not lost. While traditional perimetry is large and often immobile, head-mounted perimetry is emerging as a useful and accessible method for visual field testing, which will be accessible for spaceflight.Increased intraocular pressure and secondary glaucoma can affect peripheral vision and is one of the most common diseases of irreversible blindness on Earth. While astronauts are less likely to experience glaucoma in year-long missions, current research on goggles as a countermeasure for SANS increases pressure in the orbital area.Currently, the gold standard for testing visual field is standard automated perimetry (SAP). Used for diagnosing and monitoring glaucoma, SAP has a patient-interactive approach that helps gather perceptual deficit data in a timely, reproducible, and highly accurate manner – enabling early detection of glaucomatous changes. Some of its limitations, however, include its dependence on patient cooperation, long acquisition times, an uncomfortable posture required from patients, and prohibitive costs of equipment needed. With such limitations, it was important to explore a portable perimetry system – a characteristic of crucial importance when considering its implementation in the ISS. As the headset may be adjusted to fit neatly on the patient’s head, this perimetry device did not need a patient to maintain a specific head position, thus greatly reducing discomfort. The first renders of the virtual reality headset were not widely used due to low screen resolution and small screen sizes; however, as technology advanced, such hardware and software issues were drastically improved, resulting in a highly invaluable testing device.First described by Hollander, et al in 2000, the Kasha visual system (automated and head-mounted) yielded promising results. These significant visual changes could potentially affect an astronaut’s ability to perform critical tasks in LDSF and potentially lead to mission failure. Thus, optimizing the evaluation of such a test is of utmost importance. While the Amsler grid is available onboard the ISS, it is a highly subjective examination and various factors including the surrounding external environment may impact its ability to provide consistent data on any progression of distortion. Even small amounts of distortion detected earlier on can provide further guidance on imaging and possible interventions in the future that are approved for spaceflight.Amsler grid is onboard the ISS, which helps to identify disruptions in the macula that often manifest as metamorphopsia (distortion) of the central vision. Chorioretinal folds have previously been documented in astronauts and is one of the hallmark findings of SANS. The visual impacts of these folds are dependent on the degree of folding and proximity to the fovea, but can cause metamorphopsia or visual loss. Terrestrially, chronic chorioretinal folds lead to chorioretinal folds-related maculopathy, which leads to a decline in visual acuity.An essential component of central vision and color perception is the macula. Deficits in macular function can result in severe visual impairment, color detection, and activities of daily living in general such as reading and recognizing faces. Metamorphopsia is a very wide-spread form of visual distortion in several macular diseases . Patients with metamorphopsia often describe fluctuations in vision or deviation of straight lines. With highly sensitive precision eye-tracking, screen resolution, and wide field view capabilities, head-mounted devices are rapidly becoming an indispensable tool for visual tests.Head-mounted devices have seen a steady rise in their development and application to various terrestrial ocular diseases that include AMD, glaucoma, and amblyopia. Most recently, a study by Ong, et al in 2022 details the development of head-mounted device that could be used as a countermeasure for macular-related visual deficits. This project is supported by NASA with the goal of developing a non-invasive framework for astronaut vision.Although all of the aforementioned functional assessments of vision would be highly useful for astronauts, significant time and space limitations exist on the ISS. The addition of four additional tests of visual function may be a barrier given the current technology onboard the ISS. To address this issue, a head-mounted, multi-modal visual assessment system is currently being built to help detect the subtle visual impacts of SANS. and assess and mitigate the effects of monocular metamorphopsia from macular disruptions. This technology may one day be used in vision screening to help regions lacking adequate access to ophthalmic services on Earth.Head-mounted technology has many benefits over traditional forms of visual assessment, as it can use eye-tracking technology, with consistent illumination, be performed rapidly, and with minimal set-up required. These time-savings benefits and ease of use would help allow astronauts to perform this additional visual testing without requiring additional time from their rigorous schedules. Using this head-mounted system, previous research has been able to accurately assess DVA


Autism Spectrum Disorder (ASD) is a neurodevelopmental condition with a wide spectrum of symptoms, mainly characterized by social, communication, and cognitive impairments. Latest diagnostic criteria according to DSM-5  now include sensory issues among the four restricted/repetitive behavior features defined as “hyper- or hypo-reactivity to sensory input or unusual interest in sensory aspects of environment”. Here, we review auditory sensory alterations in patients with ASD. Considering the updated diagnostic criteria for ASD, we examined research evidence (2015–2022) of the integrity of the cognitive function in auditory-related tasks, the integrity of the peripheral auditory system, and the integrity of the central nervous system in patients diagnosed with ASD. Taking into account the different approaches and experimental study designs, we reappraise the knowledge on auditory sensory alterations and reflect on how these might be linked with behavior symptomatology in ASD. Autism spectrum disorder (ASD) is a neurodevelopment condition characterized by deficits in social communication and interaction, and restricted/repetitive behavioral features  (American Psychiatric Association (APA) In 2018, the World Health Organization updated the classification of autism in the International Classification of Diseases (ICD-11)  Over the years, the focus of ASD diagnosis was mainly related to cognitive, communication, and social impairments. But more recently, the criteria of diagnosis started to include another feature: the sensory processing domain , thus considering the recent adaption of autism diagnosis as a spectrum disorder.A.What is known about the integrity of Cognitive Function in auditory-related tasks?B.What is known about the integrity of Peripheral Auditory System in auditory-related tasks?C.What is known about the integrity of Central Auditory Nervous System in auditory-related tasks?The following questions will be addressed:This systematic review follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines  The search strategy started in September 2021 and used MeSH terms from PubMed. Used databases: PubMed, Web of Science , Scopus, and Scielo. Search terms included “Studies were eligible if they consisted of original and reported data, with comparison group design, written in English and published in peer-reviewed journals. Conference abstracts, books and documents, editorials, letters, pilot studies, case studies, reviews, and meta-analyses were excluded.Only studies with human participants with a clear diagnosis of ASD were considered. To avoid confounding results, studies with ASD participants that were also diagnosed with other symptomatology and/or disorders were excluded: epilepsy, Williams Syndrome, ADHD, Rett Syndrome, Angelman Syndrome, Fragile X Syndrome, etc. Participants diagnosed with ASD in early childhood but without current ASD symptoms, or participants with the diagnosis based on the DSM-IV without other assessments of diagnosis validation  were also excluded. No restrictions of age, gender, racial, ethnic, and socioeconomic groups were made.Studies that did not include relevant outcome data were excluded .Most of the tools used to confirm ASD diagnosis in the studies included on this review were: Short-Sensory Profile (SPP) Dunn , ChildhoCognitive function can be assessed by mental processes such as perception, attention, memory, decision-making, or language comprehension, and in the domain of social cognition . People with ASD have a profile of cognitive strengths and weaknesses, demonstrated by different levels of deficits in social and non-social cognition patterns. To understand how auditory input might influence cognitive function in ASD, it is useful to assess cognitive integrity at several domains, while performing auditory-related tasks.Communication in everyday life depends crucially on the ability to detect stimuli and dynamically shift attention between competing auditory streams. We use attention to direct our perceptual systems toward certain stimuli for further processing. Difficulties in attention processes can directly affect social interaction and communication abilities. When compared to neurotypical controls, individuals with ASD seem to have poorer performance in many auditory attention tasks: divided attention (ability to attend to two or more stimuli at the same time), sustained attention (ability to attend stimulus over longer periods), selective attention (ability to ignore details of stimulus not attended), and spatial attention . Main findings are summarized in Table Low-level auditory discrimination ability seems to vary widely within ASD. When different paradigms were compared, no differences were found for adults with ASD regarding selective attention (maintain vs switch tasks) . A study tested whether impairments in joint attention were influenced by self-relevant processing. Results showed that gaze-triggered attention is influenced by self-relevant processing and symptom severity in ASD individuals, with reduced cueing effect to voice compared to tone targets   adding different levels of task difficulty (easier and harder)   to measure brain activity in real time. EEG signals reflect electrical activity produced by the brain (brain waves). An approach known as event-related potential (ERP), uses EEG activity that is time-locked with ongoing sensory, motor, or cognitive events, to help identify and classify perceptual, memory, and linguistic operations  and a number indicating their latency in milliseconds   studies show preserved processing of musical cues in ASD individuals, but with prosodic impairments  show different P3a depending on the IQ level of ASD children. A study with high-functioning ASD children shows diminished amplitude of P3a    measures electrical signals associated with the propagation of sound information through the auditory nerve to higher auditory centers, after an acoustic stimulus. Major alterations in neuronal firing along this pathway can be detected as changes in auditory brain response . CompleMost ABR studies report differences in auditory brainstem processing for ASD individuals when compared with control groups Table . ResultsIn the first 10 ms after a click, click-ABR produce five-to-seven waveforms (wave I–VII). These wave peaks reflect the propagation of electrical activity as it travels along the auditory pathway, providing information in terms of latency (speed of transmission), amplitude of the peaks (interpreted as the number of neurons firing), inter-peak latency (time between peaks), and interaural latency (correlation between left and right ear) . Peaks can reflect either a change in the stimulus  or the periodicity of the stimulus. There are two main components in speech-ABR: the onset response (waves V and A) and the frequency-following response . The wave C represents the consonant–vowel transition, while the wave O represents the end of the vowel. Analysis of FFR includes measurements of response timing (peaks), magnitude (robustness of encoding of specific frequencies), and fidelity . Evoked otoacoustic emissions are produced by healthy ears in response to an acoustic stimulus delivered into a sealed ear canal. The acoustic stimulus causes basilar membrane motion which triggers an electromechanical amplification process by the cochlear outer hair cells, producing a sound that echoes back into the middle ear (otoacoustic emissions). These nearly inaudible emissions are measured using a sensitive microphone and help evaluate normal cochlear function. A study tested a group of children and adolescents with ASD  with normal audiometric thresholds . Integrity of the central nervous system can be assessed experimentally by several techniques, such as magnetoencephalography  is an imaging technique that is used to assess the anatomy and physiology of brain circuits. Magnetoencephalography (MEG) is a functional neuroimaging technique that detects, records, and analyzes the magnetic fields produced by electrical currents occurring naturally in the brain Cohen . While EResults from studies with high-functioning ASD individuals show delayed latencies at M50 and M100 auditory evoked responses, suggesting impairments at early auditory processes in the primary and secondary auditory cortex  seem to present greater latencies delays in M50 and M100 (components associated with language and communication skills) compared to ASD children without intellectual disabilities , theta , alpha , beta , and gamma   neurotransmission. This E/I balance seems to be very important for cortical gamma-band activity, given that gamma waves are generated through connections between GABAergic inhibitory interneurons and excitatory pyramidal cells  is a baseline shift in the electrical and magnetic signals upon exposure to a sound lasting for several seconds. The SF adapts to the probability of a sound pattern, reflecting the integration of pitch information across frequencies within the tonotopic map of the primary auditory cortex (A1)  who later developed ASD, at 3 years old . Results show dynamic functional brain networks altered in children diagnosed with ASD, including cognitive control, subcortical, auditory, visual, bilateral limbic, and default-mode network  Low-level auditory stimuli  deficits vary widely within ASD, with some interesting differences when low- and high-functioning children (based on IQ assessment) are compared Table . ASD indDifficulties in understanding others are a core feature of autism spectrum disorders  tend to show longer latencies in children with ASD, and several EEG studies show unstable neural tracking in subcortical auditory processing. However, not all children have the same clinical presentation, suggesting the potential existence of auditory processing-related subtypes of children with ASD. Considering the results from ABR studies, one interesting finding is ABR longitudinal profile across development that seems to indicate shorter latencies in newborns, longer latencies in ASD children compared to newborns, no differences in adolescents, and normal ABR in adults. Infants and toddlers later diagnosed with ASD present delayed auditory brainstem responses, mainly with later wave V latencies  (Stevenson et al. The usefulness of longitudinal EEG as a diagnostic tool for uncovering developmental trajectories in ASD has been recently suggested, showing that delta and gamma frequency power trajectories can differentiate autism outcomes, in the first postnatal year (Gabard-Durnam et al. Not many studies have looked into E/I balance related to auditory processing in ASD. The imbalance hypothesis is highly promising, given the suggestion of cortical excitatory–inhibitory imbalance in areas related to auditory processing in subjects with ASD (Murray et al.


Upon their derivation from human pluripotent stem cells (hPSCs), organoids reveal the blueprints of human organogenesis, further allowing the faithful recapitulation of their physiology. Nevertheless, along with the evolution of this field, advanced research exposed the organoids’ shortcomings, particularly regarding poor reproducibility rates and overall immatureness. To resolve these challenges, many studies have started to underscore the relevance of mechanical cues as a relevant source to induce and externally control hPSCs differentiation. Indeed, established organoid generation protocols from hPSCs have mainly relyed on the biochemical induction of fundamental signalling pathways present during kidney formation in mammals, whereas mechanical cues have largely been unexplored. This review aims to discuss the pertinence of (bio) physical cues within hPSCs-derived organoid cultures, while deciphering their effect on morphogenesis. Moreover, we will explore state-of-the-art mechanobiology techniques as revolutionizing means for understanding the underlying role of mechanical forces in biological processes in organoid model systems.Recently, organoids have emerged as revolutionizing tools with the unprecedented potential to recreate organ-specific microanatomy  The kidney plays a vital role in regulating bodily fluids and maintaining their composition. It performs crucial tasks such as blood filtration, hormone regulation, and waste removal, all of which are essential for maintaining organism homeostasis. The kidney’s remarkable anatomy enables it to carry out these functions effectively: its highly vascularized structure allows for efficient filtration from the blood vessels, while its connection to the urinary tract facilitates the excretion of waste in the form of urine. These processes are carefully regulated by signalling hormones within the body, which are essential for maintaining blood pressure, electrolyte concentration and acid-base balance.The kidneys contain approximately 1 million nephrons, which are the functional units . Each neIn humans, the formation of nephrons (nephrogenesis), is completed around 36 weeks of pregnancy, ensuring the kidney’s complexity is established by birth. Throughout this developmental time, various environmental cues and genetic factors guide the precise localization and formation of tissue components within the organ. While the genetic and biochemical cues involved in kidney development are well understood, the role of mechanical cues has remained largely unexplored.in vivo remain unknown.Given the kidney’s intrinsic complexity and its vital role in maintaining overall organism homeostasis, it has gathered significant interest in the scientific community. Researchers have been particularly focused on unravelling the morphogenetic principles that govern kidney development from early embryonic stages until birth. Advancements in developmental studies, including the use of human pluripotent stem cells- (hPSCs) or adult stem cell-derived organoids, have pushed the boundaries of our understanding of organogenesis and disease progression in this complex organ. However, despite these major breakthroughs, the precise mechanisms underlying the (bio) mechanical machinery that contributes to renal fate specification and the overall tissue organization As the recognition of mechanical cues in development grows, there is an increasing demand for sophisticated bioengineering solutions to understand the mechanical signals and responses involved in organogenesis. Mechanobiological techniques, as reviewed in , can helin vitro kidney organoid differentiation, which pose as valuable tools for studying development and disease. The review will also discuss what is known so far regarding the presence of mechanical cues during embryo and organ development and will highlight state-of-the-art techniques commonly employed in the field of tissue mechanics to probe cell-to-tissues responses. At the same time, here we aim to provide a comprehensive view on how these tools may be translated to understand kidney morphogenesis exploiting hPSCs-derived kidney organoid technology. Finally, the review will explore the rising need for more interdisciplinarity between the fields of classical developmental biology and mechanobiology, which can significantly enhance our understanding of organ development and disease progression in humans.This review aims to summarize current knowledge on mammalian kidney development and provide insights into the state-of-the-art methodologies for The development of the adult mammalian kidney begins during embryogenesis with the formation of three precursor structures: pronephros, mesonephros and metanephros. The prior two are transient, yet still necessary structures, the absence of which have been associated with renal agenesis. The metanephros on the other hand is the permanent structure that gives rise to the functional adult kidney . PrimitiThese RV structures elongate into polarized comma-shaped bodies, then S-shaped bodies, readily connected to the UB-derived collecting duct. S-shaped bodies reportedly evolve into mature nephrons, exhibiting glomerulus and tubular structures, including the proximal tubule, loop of Henle and distal tubule. All these structural changes are known to be guided by the genetic blueprint. In humans nephrogenesis repeats until shortly before birth.Like any other developmental process, kidney organogenesis exhibits a remarkable sensitivity. Indeed, disruptions or impairment at any stage of kidney morphogenesis can precipitate a cascade of severe inborn conditions, ultimately known as congenital anomalies of the kidneys and urinary tract (CAKUT). As such, CAKUT encompasses a family of urinary system malformation at birth, including kidney defects such as renal agenesis or hypoplasia, as well as ureteric malfunctions . These aSeveral CAKUT manifestations have been reported to be clinically associated with hypertension, ultimately leading to, in certain occasions, end stage renal disease (ESRD). There is therefore an urge to understand the mechanisms underlying CAKUT anomalies in the human context. Indeed, several works have been developed in knock-out animal models, however, the phenotypical manifestation of the pathological mutations may vary from species to species .Mammalian kidney development is a fascinating process that exhibits remarkable evolutionary conservation across species. The study of human-specific kidney development throughout gestation has been challenged by limited access to human embryonic kidney samples and the ethical concerns surrounding their origin. However, this limitation has been overcome by utilizing animal models of lower complexity, such as rats and mice, to investigate the intricate cellular and molecular processes involved in kidney organogenesis. By leveraging the remarkable conservation of embryogenesis across mammalian species, findings from these animal models have provided valuable insights into human kidney development . In factComparison of embryonic kidney development across diverse mammalian species has revealed high levels of similarity. Notably, studies have documented shared morphological features between mouse and human embryonic and adult tissues, demonstrating clear parallels in tissue architecture at species-specific gestational timepoints . The bluDespite the diversity of mammalian species, the fundamental principles and mechanisms governing kidney development remain remarkably similar. Indeed, our current understanding of kidney organogenesis has been derived from mouse and rat embryonic kidney samples. Shared anatomy, patterning and signalling pathways have emphasized the utility of mouse embryonic models in recapitulating kidney development when human samples are unavailable. And while the overall process of embryonic kidney development remains evolutionarily conserved among mammalian species, inter-species molecular and anatomical variations still exist . Indeed,With regards to inborn conditions affecting the kidney, the mechanisms underlying CAKUT, for instance, have been more challenging to address in non-human models. Importantly, heterozygous or homozygous mutations in mice models have caused either embryonic or postnatal lethality, making it difficult to study the underlying pathophysiology .in vivo and in vitro) allows for the interrogation of early stages of cell lineage specification and differentiation. Thanks to this, specific cell culture conditions sustaining and promoting cell differentiation have been identified and are now consistently used as a tool in developmental research. For more than 2 decades researchers world-wide have started to define external conditions aiming to guide hPSCs differentiation while exploiting inherent characteristics of these cells, that are capable of self-organization and symmetry breaking  has significantly impacted how scientists study human development and diseases outside of the human body . The capbreaking . While ibreaking , seminalbreaking .In vivo, different cell types arise from stem cells through a process of lineage commitment and cell sorting, that allow a spatial organization like an organ during development. In vitro, self-organized collection of hPSCs-derived cell types can recapitulate, until certain extent, specific functions of an organ.A technical evolution of EB culture systems led to the overall explosion of the organoid field from hPSCs. This was based on the seminal work from Yoshiki Sasai on the establishment of the first self-patterned stratified cortical tissues generated by plating EBs in serum-free medium on a coated surface . At the in vitro renal inductive signals that occur during kidney organogenesis  and polycystic kidney diseases (PKD), among others [reviewed in more detail in ]. While PAX2, HNF1B, BMP7, RET, GATA3, SALL1, SIX5, and EYA1 among others developmental genes have been previously reported.In this regard, CAKUT is an umbrella term encompassing an extensive spectrum of embryonic kidney and urinary tract malformations that affect approximately 1 in 100–500 new-borns . The phePAX2) is an important developmental gene in kidney organogenesis, expressed throughout nephron differentiation: starting from progenitors to epithelial renal vesicles to distal fragments of the renal tubules. Particularly, in mice, Pax2 was shown to play an indispensable role in MET of NPs, and the positioning and outgrowth of the ureteric bud  has been shown to play an important role in nephrogenesis. Studies in zebrafish have identified the role of this transcription factor in nephron patterning and segmentation, where embryos lacking hnf1ba/b would not express markers characteristic for distal and proximal segments, yet still form an epithelial tubule  can be roughly identified during the differentiation protocol  and create connections like cell-to-ECM and cell-to-cell junctions that are necessary for transmitting the biomechanical signals towards the nucleus and ultimately affect cell function. This ability of cells to sense their mechanical microenvironment is termed in utero, the embryonic microenvironment contains a plethora of ECM proteins  and a highly viscoelastic matrix (ranging from 0.1 to 100 kPa in stiffness) which has been shown to influence the fate of stem cells within the forming tissue . Studies have shown how upon exposure to soft matrices, these multipotent MSCs can be driven to the adipogenic lineage, while stiffer microenvironments can induce osteogenesis in the same naïve stem cells , polydimethylsiloxane (PDMS), and polyethylene glycol (PEG), among others. In this section the use of natural and synthetic substrates for organoid generation and disease modelling applications will be revisited.in vivo, making it a perfect candidate for long term maturation of hPSC-derived organoids in vitro [reviewed in detail in (Decellularized ECM (dECM) has the advantage of maintaining the natural architectural composition of tissues etail in ]. Dependetail in . HoweverNaturally occurring ECM proteins, such as collagen or laminin can also be used to generate scaffolds for tissue engineering purposes [reviewed in ]. CollagOrganoids have also been grown in natural polysaccharides like alginate, an inexpensive, biocompatible and readily-tuneable substrate suitable for biological application [reviewed in ]. SeveraSynthetic polymers, on the other hand, allow a precise control over the mechanical, chemical and structural elements of the substrate. By adjusting the composition of synthetic substrates, it becomes possible to mimic the viscoelasticity, porosity and density of certain tissues and study the effects of mechanical and chemical cues on cell fate decisions.in vitro tissues (Polyethylene glycol (PEG) is a nontoxic and biocompatible polymer, increasingly utilized for cell culture and controlled release of biomolecules within  tissues  [see in  tissues ]. It is  tissues .Polyacrylamide (PAA) hydrogels can be tuned to a wide variety of stiffnesses (ranging from 0.2 kPa up to 200 kPa) depending on the compositional ratio of acrylamide and bis-acrylamide. In 1997, using this synthetic hydrogel, it was demonstrated for the first time that substrate stiffness can indeed affect cell adhesion, spreading and migration . FollowiPolydimethylsiloxane (PDMS), a material primarily intended for microfluidic applications  has beenin vitro culture conditions to natural in vivo tissue microenvironments. Moreover, since synthetic tuneable materials allow a tight control of substrate stiffness, porosity, density, etc., it becomes possible to perturb the mechanobiological environment of cells and question the way tissues respond to mechanical input from their environment.Ultimately, the use of such versatile biocompatible natural and synthetic materials allows a much closer resemblance of Thus far, many studies have evidently demonstrated how mechanical stimuli like stiffness, viscoelasticity and fluid flow can influence cell fate decisions , and havin vivo, in vitro, and ex vivo can provide valuable insights into the underlying mechanisms of these processes.Mechanical forces are involved in various biological processes, including cell division, tissue growth, and organ function. The ability to measure these forces in vitro (most)   , micropillars, microdroplets/micro-inserts, cantilevers/atomic force microscopy (AFM). These techniques can be reliably used to measure cellular forces o (most) , ex vivovo (AFM) , and everoplets) .Other techniques, like FRET tension based sensors, rely on molecular linkers/fluorophores that can allow the measurement of tensile forces between cells and their extracellular environment  and evenxenopus, zebrafish) at early stages of development have been widely explored with the use of micropipette aspiration and laser ablation techniques  .When cells are exposed to an external mechanical stimuli, the first line players that initiate the mechanotransduction pathway are mechanosensitive membrane proteins . These iThe high sensitivity of these membrane proteins to instigate mechanotransduction further down the cell, makes it a potent candidate for tissue engineering purposes. By regulating such ion channels through, for example, genetically induced light activation, it could become possible to control cellular responses in the absence of mechanical stimuli.Further down the mechanotransduction pathway are mechanosensitive transcription factors that inititate the activation of genes in the nucleus to prompt a cellular response to mechanical stimuli.Yes associated protein (YAP)/Transcriptional coactivator with PDZ-binding motif (TAZ) are one of the most known mechanosensitive transcriptional regulators, which evidently play an important role in organogenesis . They arIn the kidneys, TAZ activity has been shown to be associated with cystogenesis in PKD, implying the important effect of mechanotransduction during kidney disease progression .At the level of stem cells, depletion of this key transcription factor (YAP) has been shown to result in a loss of pluripotency, while constituent expression leads to maintenance of stemness . StudiesThese studies demonstrate the importance of understanding the mechanosensing and mechanotransduction machinery in cells as a bottom-up approach to control morphogenetic events during development. Here the use of human PSC-derived organoids can serve as a framework to question the mechanical aspects of organ development, where bioengineering advances can aid in the realization of these studies in a controlled and reproducible manner.in vivo, researchers gained an in-depth knowledge regarding the principles governing nephrogenesis, an invaluable asset to ultimately recreate this process in vitro. While in vitro models of human kidney organogenesis can to some extent faithfully recapitulate the organ’s microanatomy and physiology, there are still major limitations in reproducibility, control of cell type, composition, and lack of vascularized and innervated structures. Here, by acknowledging the mechanical microenvironment of in vivo tissues during development, and by closely mimicking these external cues, it becomes possible to drive organoid maturation a step further and gain a better understanding of organogenesis and disease progression.Altogether, accumulated findings within the last decades have revealed the immense value of hPSC-derived models as a platform on which organogenesis could be recapitulated. By investigating the specification of stem cells towards the kidney lineage in vitro, there is still a long way to go. In order to guide cells in the direction of organ development, biochemical and mechanical signalling cascades within and between cells need to be perfectly coordinated to navigate through the complex dynamics of tissue patterning. Hence in kidney organogenesis, more consistent efforts are needed to truly understand the underlying biomechanical machinery and how it works in concert with the genetic program. Only then will it be possible to engineer functional kidney tissues.While current advances in bioengineering and mechanobiology are paving new strategies for studying and guiding tissue morphogenesis


Amyloids are high-order proteinaceous formations deposited in both intra- and extracellular spaces. These aggregates have tendencies to deregulate cellular physiology in multiple ways; for example, altered metabolism, mitochondrial dysfunctions, immune modulation, etc. When amyloids are formed in brain tissues, the endpoint often is death of neurons. However, interesting but least understood is a close connection of amyloids with another set of conditions in which brain cells proliferate at an extraordinary rate and form tumor inside brain. Glioblastoma is one such condition. Increasing number of evidence indicate a possible link between amyloid formation and depositions in brain tumors. Several proteins associated with cell cycle regulation and apoptotic pathways themselves have shown to possess high tendencies to form amyloids. Tumor suppressor protein p53 is one prominent example that mutate, oligomerize and form amyloids leading to loss- or gain-of-functions and cause increased cell proliferation and malignancies. In this review article, we present available examples, genetic links and common pathways that indicate that possibly the two distantly placed pathways: amyloid formation and developing cancers in the brain have similarities and are mechanistically intertwined together. Brain tumor is a widely used term encompassing multiple diseases of heterogeneous nature. Malignant and benign tumors of central nervous system (CNS) include more than hundred distinct pathologies with highly diverse etiology and epidemiology. Despite being less common, brain-associated cancers share disproportionately higher mortality among all cancer types. According to American Cancer Society’s data, more than 300,000 cases of primary CNS  tumors have been recorded globally in 2020, while approximately 250,000 people died in same year with these tumors. In addition, incidences of secondary brain tumors are also prevalent adding up to these numbers. Secondary brain tumors originate extracranially but travel to and spread inside the brain. Five-year survival rate for brain cancer patients is 36% in United States. Contrary to extracranial tumor types, CNS or brain tumors show specific physiological characteristics owing to the microenvironment present in the brain tissues. The unique cell types, metabolic constraints, and anatomy of the brain present exceptional technological challenges in understanding and developing treatments. One specific feature that brain tumors present is the deposition of amyloid-like formations in the brain tissues harboring the tumors. In 1981, Spaar et al. reported amyloid-like structures in surgically removed brain tumor mass from a forty-six-year-old woman [Amyloids are one of the most highly studied proteinaceous assemblies, yet our understanding of these structures is not fully grown. In general, several proteins (or peptides) can enter into an amyloid-like state unless prevented by molecular chaperones . These pThe most common tumor suppressor protein TP53 (p53) has been observed to self-polymerize and accumulate inside the breast and skin cancer cells . As showA line of accessory factors, other than amyloid deposits, show common origin and progression of the two disease conditions  in the brain. For example, presence of a conducive environment in and around the brain tumors can affect multiple proteostasis pathways affecting proteome-wide alterations leading to protein aggregation and amyloid deposition. Several common environmental risk factors that include but are not limited to metal ions like mercury, lead and arsenic; electromagnetic fields; smoking, hair dyes; pesticides; benzene; petroleum products and lubricating oil, etc., also harbor both types of pathological changes . In addiIn this article, we are presenting a brief literature on the available common links among the conditions that include, but are not limited to protein aggregation, amyloid formation, brain tumor malignancies and proliferation. We have taken glioma and AD as two most studied cases of the two classes of the disease in question.Tumors are abnormally grown non-functional tissue mass primarily caused due to excessively high rates of cell proliferation. These atypical growths could be both benign or malignant and may eventually turn out to be a pathological lump leading to a number of diseases, including cancers of different organs or tissues. The definition of a primary brain tumor appears very simple as a tumor originating from the brain; however, the complexity is much higher than many other cancer types. Based on the site of origin and histopathology, primary brain tumors may fall into the category of glioma, schwannomas, glioblastoma, pineal region tumor, meningioma, pituitary adenomas, and a few other kinds of tumors ,22. The Among these mutated proteins IDH plays a key role in maintaining cellular homeostasis in tumor cells by catalyzing oxidative decarboxylation of isocitrate in Kreb cycle. Further, the metabolic reprogramming by IDH mutation generates redox imbalance and epigenetic alterations, making this gene a prominent therapeutic target ,30. PTENBrain tumor diagnosis mostly relies on magnetic resonance imaging or positron emission tomography . The curDementia is one of the common symptoms among brain tumor patients. Moreover, surgical interventions also impact the memory of brain tumor patients . A case Malignant brain tumors are well-known for deaths of neurons. The mechanism of neuronal cell death in brain tumors is similar to neurodegenerative disorders, like Alzheimer's disease . It is aInactivation of tumor suppressor p53 gene due to mutations is one common factor found in ∼50% glioma patients. Previous studies showed positive impact of virus-mediated p53 gene therapies by markedly reducing the growth of glioma and prolonging the lifespan ,72. In aSeveral tumor types have been found associated with local (within tumor) or systemic (elsewhere) deposition of amyloid aggregates. For example, occurrence of amyloid aggregates of transthyretin, immunoglobulin light-chain, and amyloid beta in breast cancer tissues has been reported in several previous studies . AmyloidIn 1981, Spaar et al. reported deposition of walnut-sized amyloid mass in a patient suffering from progressive visual loss with scotoma and finally from hemianopia, associated with attacks of headaches and recurrent episodes of depression . PrimaryOne most relevant example connecting these two conditions: the cancer and amyloids is the tumor suppressor protein TP53. This p53 protein has been extensively studied for its roles in cell cycle regulation. Mutation in p53 gene is a common factor involved in pathological manifestation of plethora of cancers. For example, M237I mutant of p53 confers chemoresistance to a highly potent chemotherapeutic drug temozolomide in glioblastoma cell lines . InitialGlioblastoma is often referred as a cold-tumor due to presentation of an immunosuppressive environment. Although the disease can occur in any age group, its higher prevalence in late-age individuals is very common. Aging brain consists of a number of pro-inflammatory changes primarily mediated by several age-associated genetic manipulations happening in the brain . SerpinaTriggering receptor expressed on myeloid cells-2 (TREM2) is an innate immune receptor which is expressed on the cell membrane of monocyte-derived dendritic cells, osteoclasts, and microglia ,129. TRETREM2 role has also been suggested to increase with AD-related neurofibrillary pathology in a study conducted on postmortem inferior temporal cortex samples obtained from 60 patients . These p+ T cells and thus, the overall tumor growth [+ tumor associated lymphocytes in tumor microenvironment [–/– and wildtype mice in 3-methylcholanthrene-induced sarcoma, colorectal cancer and mammary tumor showed that anti-PD-1 immunotherapy is more effective in Trem2 deficient mice as compared to wild type mice. Anti-TREM2 monoclonal antibody treatment prior to PD-1 immunotherapy that made tumors more susceptible and increase the efficacy of the former drug in different cancer types, including glioblastoma [TREM2 have different roles in various pathologies, cancer and cell types. For example, substantial evidence confirm a prominent immunosuppressive impact of TREM2 in cancer cells. The cell-specific TREM2 functions, such as in fibroblast and stromal cells are different than the actual tumor microenvironment . Deficier growth . On the ironment . Anotherblastoma ,143. It Neuroinflammation is defined as a complex inflammatory response in the brain and other parts of the central nervous system in response to injury, infection, stress, neurogenerative diseases, age, and cancer . ReleaseAs mentioned, brain tumors are primarily known for their immune-suppressive tumor microenvironment; but generate a neuroinflammatory response within tumors that supports tumor metastasis and invasion ,154. Neuvice versa. Several common factors, including environmental, and genetics have been identified so far that can induce one of the two and end up in exaggerated deterioration caused by the other pathway complementing the first one. Recent studies suggest that plaque surrounding microglia are highly specialized cells, harboring a distinct subset of proteome compared to other microglial cells. These specialized cells try to combat the spread of the plaque by inducing damage-sensing pathway gene networks. In addition, the cells fine-tune their housekeeping and tolerogenic functions in order to slow down the loss of neurons by eating more of defective molecules [In his initial drawings of affected AD brain regions, Dr. Alois Alzheimer has depicted glial cells huddle around the degenerating neurons. Now, after a century of research, we know with certainty that inflammation plays very important role in the mediating neurodegeneration or olecules ,165. Treolecules . Both p5olecules . The comSeveral attempts have been made to exploit the close connections (crossovers) among the diseases belonging to the two classes: neurodegeneration and cancers. For example, some therapeutic success has been recorded in terms of overcoming cell malignancies and controlling the tumor growth when subjected to anti-amyloid compounds. Major compounds that showed tremendous success in providing therapeutic benefits in the two diseases are: resveratrol, trehalose, myricetin, quercetin, EGCG, rapamycin, etc. . MyricetDespite the link between brain malignancies and memory loss is slippery, efforts are ongoing to understand this dynamic relation between the aggregation and tumor formation. The clinical interest has gone up in recent years, but we still don't know if enhancing the proteostasis pathways or suppressing neuroinflammation would benefit in neurodegeneration. Similarly, our knowledge about modulation of proteostasis pathways and their impact on cancer progression is elusive. In past years, multiple studies showed that small anti-amyloid drug candidates show high potential for inducing apoptosis, inhibition of tumor growth and malignancies. Contrary to these examples, not many anti-cancerous drug molecules show similar effects in preventing amyloid formation and neurodegeneration. In addition, there are multiple limitations to these studies as well. First, we do not have mouse models that can truly recapitulate AD pathology. Second, it is very difficult to find right patient subjects for testing new drug candidates. In conclusion, neurodegeneration (by amyloid formation) and brain cancers (e.g. glioma) seem to be two pathological conditions that are physiologically opposite and are distantly placed on the scale of cell cycle progression and cell death. However, common molecular signatures, cellular pathways and tissue-specific changes affect both conditions and indicate a close association between two. Future studies investigating the system wide changes may provide more conclusive evidences and reconnect the missing dots.


Methods in Mammary Gland Biology and Breast Cancer was held on April 26th − 29th in Weggis, Switzerland. For the first time, early career researchers organised and took part in an additional ECR workshop on the 26th of April, which was received with great enthusiasm. The topics of the main workshop included mammary branching and morphogenesis, novel experimental systems (model organisms), systemic influences on tumour progression and the tumour microenvironment. Novel and recent findings were shared across excellent oral and poster presentations.The fourteenth annual workshop of the European Network for Breast Development and Cancer (ENBDC) on  Methods in Mammary Gland Biology and Breast Cancer in Weggis, Switzerland. The primary goal of this annual workshop is to bring together researchers in an informal setting to encourage in-depth and unvarnished discussions of novel technologies and discoveries [On April 26th -29th 2023, the European Network for Breast Development and Cancer (ENBDC) held the 14th workshop on For the first time in its history, the ENBDC workshop was preceded by a one-day workshop for early career researchers (ECRs), similar to the Gordon Research Seminars that precede every Gordon Research Conference. The inaugural ECR workshop, organised by Silke Blair Chalmers  and Tanne van der Wal , provided an opportunity for learning and networking through scientific sessions, joint discussions of technologies and protocols, and social activities. The scientific sessions consisted of engaging presentations from 9 talented postdocs and PhD students, selected by the organisers: Patrycja Szybowska , Oliver Cottrell , Aida Aide Luna Perez , Benjamin Davies , Calvin Rodrigues , Mathilde Folacci , Neve Prowting , Marleen Aarts , and Marika Caruso . The presentations were very well received with active interaction between peers and constructive discussion of methods and results. The ECRs also participated in a technical workshop to troubleshoot their work surrounding the chosen topics “imaging and image analysis” and “inducible gene expression”. The organisers of the ECR workshop are grateful to all the participants of this inaugural event and are hopeful it will become a mainstay of future ENBDC meetings.The main meeting commenced with a keynote address by Christine Watson . The address provided an opportunity for the ENBDC community to acknowledge and celebrate the tremendous contributions Watson – who recently retired – has made to the field of mammary gland biology. Watson provided a compelling overview of her career, commencing with work identifying and characterising the role of the KRAB zinc finger protein ZFP157 , in pregnancy and lactation –6. Loss Next, Watson outlined work exploring the role of mammary stem cells in mammary gland development . ThroughSwitching to involution, Watson discussed the characterisation of the cell death pathways in post-lactation mammary gland regression –11. OrigIn the final part of her keynote lecture, Watson discussed recent work exploring the immune environment of the mammary gland . They foThe first scientific session on “Dimensions of development, branching and morphogenesis” was chaired by Walid Khaled . Biophysicist Edouard Hannezo  opened the session presenting insightful work exploring self-organized branching morphogenesis. Although branching morphogenesis occurs ubiquitously in glandular epithelia, the rules governing this process are not fully understood . To explWnt gene was discovered as a mammary oncogene in mice. As small aberrations in WNT signalling might contribute to mammary tumorigenesis, van Amerongen decided to first clarify the tissue-specific expression of WNTs in the mammary gland, focussing on WNT4, which is known to be downstream of progesterone signalling [Wnt4 promoter. Interestingly, CRS4 itself seems to function as a hub for other enhancers, including for progesterone receptor (PR) dependent enhancers. Moreover, luminal specific transcription factors, including GRHL2, were discovered as critical Wnt4 regulators.Next, Renée van Amerongen  presented her unpublished work on WNT4 regulation in the mammary gland, starting with the important question of tissue-specific expression patterns. Although WNT signalling is a well-known oncogenic pathway in particular in colorectal cancer, its precise role in breast cancer remains undetermined, even though the first gnalling . In bothThe second session, focussing on “Novel models and methods for mammary gland studies” was chaired by Colinda Scheele . The session was commenced with an insightful presentation from Kate Hughes , who divided her time between providing an overview of the comparative physiology of non-traditional models and describing her research in mammary gland development in ruminants. While acknowledging the central role of laboratory rodents in mammary gland research, Hughes highlighted how non-traditional animal models may provide additional insights into mammary gland biology due to similarities in their biology and histo-anatomy . In concSandra Schöniger  gave the final presentation of the session, focussed on the characterisation of mammary tumours in pet rabbits. Rabbits may represent a novel model to study breast cancer, as they develop spontaneous mammary tumours, have a life expectancy of 6–13 years and relatively low housing costs. Other supporting factors are very similar histological features between mammary glands in humans and rabbits, and that the size of some rabbits that can allow repeated biopsy collections, e.g., to check effectiveness of cancer therapies . PredictSession 3, “Systemic influences on tumour progression and therapy response” was chaired by Jos Jonkers  and kicked off by Emmanuelle Charafe-Jauffret , who provided an in-depth overview of her work on tumour initiation and breast cancer heterogeneity , 25. Pre+ T cells, which in turn increased eosinophil production in the bone marrow. Furthermore, mice treated with ICB and cisplatin had improved responses in comparison to ICB treatment alone, which Blomberg discovered was due to increased IL33 levels promoting intra-tumoral eosinophil infiltration and subsequent activation of CD8+ T cells. Collectively, this study provided evidence of an IL5-IL33-eosinophil axis which promotes ICB response, and opens the possibility of future approaches utilising this axis to improve patient outcomes.The second invited speaker was Olga Blomberg , who presented her work on the role of eosinophils in the response to immune checkpoint blockage (ICB) in breast cancer . AlthougBRCA1/2 mutation carriers had gene signatures indicative of immune exhaustion, which may suggest immune escape mechanisms are present in very early stages of tumour formation. Furthermore, there were distinct differences in BRCA1 carriers in comparison to BRCA2 carriers. This atlas, which has already provided novel insights into cellular heterogeneity, will be a tremendous resource for the community.The last invited speaker in this session was Walid Khaled , who graciously replaced a last-minute apology to present recent work exploring tumour initiation via single-cell analyses in human samples . UnderstThe session then progressed to the first selected short talk for the meeting from Zuzana Sumbalova Koledova , who discussed recent work exploring the stromal environment during mammary gland morphogenesis via spatial transcriptome mapping. Although the stromal environment has been long recognised to play a key role in mammary gland branching morphogenesis, its molecular function and the potential existence of specialised stromal cells is still unknown. To address this, Sumbalova Koledova and her team conducted single-cell RNA sequencing on pubertal mammary stroma combined with spatial transcriptome mapping. Using this approach, they identified specialised, spatiotemporally restricted fibroblast subpopulations that were unique to pubertal mammary glands and coincided with the presence of terminal end buds. Extending this work by lineage tracing as well as in vitro studies in organoid co-cultures, they demonstrated the functional heterogeneity of fibroblast subpopulations during mammary morphogenesis. This work provides an extensive functional classification of fibroblasts roles in the developing pubertal mammary gland and will have implications for the understanding of our stromal involvement in other settings, including pathologies such as fibrosis or cancer.G12D-RFP or PIK3CAH1047R-RFP) marking mutant cells, or fluorescent reporter only  marking wild-type cells. Combining this model with lineage tracing and together with imaging, Ciwinska found that oncogenic expression in the mammary impacted the growth and survival of both, oncogene-expressing mutant cells and surrounding wild-type cells. She demonstrated that the impact on wild-type cells was cell lineage dependent, with wild-type basal and luminal cells responding differently to the presence of surrounding red labelled mutant cells. Moreover, the behaviour of mutant cells was found to differ depending on the oncogene expressed. This work provides novel insights into the dramatic morphological remodelling in the mammary gland following the emergence of pre-cancerous mutant cells.The final short talk of the session was provided by Marta Ciwinska , who presented her work exploring alterations in mammary gland morphology upon oncogenic expression. Initial stages of breast cancer development, including how mutant cells spread and proliferate, and the responses of healthy tissue to mutant cells, are poorly understood. To explore this, Ciwinska utilised an inducible multi-colour reporter mouse model that recombined to stochastically express either a fluorescent reporter and an oncogene  chaired the fourth and final session of the meeting, focussed on “The mammary microenvironment and tumour progression”. The session was commenced by Ilaria Malanchi , who presented her recent work on metastatic dormancy and reactivation. In patients, disseminated tumor cells (DTCs) are often maintained in the tissue in a state of dormancy for many years prior to outgrowth. What causes DTC reactivation is largely unknown. Malanchi presented an in vivo model of dormancy-permissive tissue where metastatic cells are largely kept dormant to investigate what perturbations cause metastatic reactivation.Clare Isacke  gave an excellent overview of her work on the tumour microenvironment in estrogen receptor-positive (ER+) breast cancer . PatientThe first short talk of this session was provided by Amelia Hasenauer , presenting her ground-breaking engineering work on 3D bioprinting of mammary gland tissue using decellularized matrix materials. She presented her workflow of creating biomaterial scaffolds by decellularization of the bovine mammary gland, producing material with a gelling temperature below 37 °C. By mixing this matrix with gelatine and a photo crosslinker she was able to produce a photoreactive biomaterial to use for 3D bioprinting, allowing flexibility in creating mammary gland-like shapes and extrusions. She showed adhesion of both MCF10A cells and primary bovine mammary epithelial cells to 3D printed biomaterials in a variety of shapes.The final short talk of the meeting was presented by Qiang Lan , who discussed his pioneering work on epithelial-mesenchymal interactions during embryonic mammary gland development, focussing on the initial branching events of the embryonic mammary gland epithelium . Lan shoThe DeOme Award for the best short presentation was given to Zuzana Sumbalova Koledova . In another first for the meeting, the poster prizes were renamed to pay tribute to researchers who have contributed tremendously both to their respective fields, as well and to ENBDC community. The Glukhova-Medina poster prize, named after Marina Glukhova and Dan Medina, was awarded to Oliver Cottrell  for his work on the oscillatory HES1 expression in MCF7 cell cycle progression and arrest. Calvin Rodrigues , was awarded the Hynes-Streuli poster prize, which honours Nancy Hynes and Charles Streuli, for his work on the transcriptional responses induced by Notch activation in vivo in mouse basal mammary cells. Finally, the Watson-Rosen poster prize, celebrating Christine Watson and Jeffrey Rosen, was awarded to Larissa Mourao , for her work on developing 3D human breast organoids to explore step-wise developmental stages during tumour initiation.The broad range of topics and exciting new research presented during the 14th ENBDC meeting was met with great enthusiasm from all participants. It was decided unanimously that the ECR workshop was a success and would be held again. The 15th ENBDC will be held on May 1st – 4th 2024 and will include an ECR workshop on May 1st, to which all interested early career researchers in the field are most welcome and encouraged to attend. The meeting will be chaired by Silvia Fre , and co-chaired by Walid Khaled . The postdoc and PhD chairs will be Beata Kaczyńska , Maria Rafaeva  and Jaime Redondo .


Clinicians should be aware that the immune microenvironment not only influences biological characteristics of HCC but also provides a reliable theoretical basis for comprehensively evaluating tumor responses after immunotherapy. Combining multi-parameter MRI with radiomics and deep learning can help to explore the biological characteristics of HCC. Future artificial intelligence technologies may extract biologically interpretable multidimensional structures from MRI images more effectively, thereby promoting precise and visual classification of HCC immune microenvironments. This advancement of MRI combined with multi-omics could potentially assist in individualized treatment planning.In recent years, significant advancements in immunotherapy for hepatocellular carcinoma (HCC) have shown the potential to further improve the prognosis of patients with advanced HCC. However, in clinical practice, there is still a lack of effective biomarkers for identifying the patient who would benefit from immunotherapy and predicting the tumor response to immunotherapy. The immune microenvironment of HCC plays a crucial role in tumor development and drug responses. However, due to the complexity of immune microenvironment, currently, no single pathological or molecular biomarker can effectively predict tumor responses to immunotherapy. Magnetic resonance imaging (MRI) images provide rich biological information; existing studies suggest the feasibility of using MRI to assess the immune microenvironment of HCC and predict tumor responses to immunotherapy. Nevertheless, there are limitations, such as the suboptimal performance of conventional MRI sequences, incomplete feature extraction in previous deep learning methods, and limited interpretability. Further study needs to combine qualitative features, quantitative parameters, multi-omics characteristics related to the HCC immune microenvironment, and various deep learning techniques in multi-center research cohorts. Subsequently, efforts should also be undertaken to construct and validate a visual predictive tool of tumor response, and assess its predictive value for patient survival benefits. Additionally, future research endeavors must aim to provide an accurate, efficient, non-invasive, and highly interpretable method for predicting the effectiveness of immune therapy. Primary liver cancer (PLC) is the fourth most common malignant tumor worldwide and the second leading cause of cancer-related deaths. PLC poses a significant threat to people’s lives and health. Approximately 75% of PLC cases are attributed as hepatocellular carcinoma (HCC) . Due to The development and progression of HCC involve a multifactorial and multistep process, including interactions between hepatitis B virus, hepatitis C virus infection, and chronic liver inflammation response . IncreasThe PD-1/PD-L1 and CTLA-4 immune checkpoint pathways within the immune microenvironment of HCC are among the important mechanisms by which tumors induce immune escape . This meSubsequently, although a series of randomized, open-label phase II/III clinical studies have reported favorable overall survival and progression-free survival benefits of immune checkpoint inhibitors (ICIs) in patients with advanced HCC, the objective response rate of the lesions is still at a low level (approximately 25%) ,9. StudiThe current expert consensus suggests that due to the relatively short clinical application time of immunotherapy for HCC, there is still a lack of experience in various aspects, including patient selection, treatment regimen determination, and efficacy assessment . FurtherTo gain a deeper understanding of the potential impact of different states of the tumor immune microenvironment on tumor development and the response to tumor immunotherapyin hepatocellular carcinoma (HCC), some researchers have classified the tumor immune microenvironment based on relevant information about the enrichment of major immune cells. Kurebayashi et al.  classifiIn addition, the 2020 European Association for the Study of the Liver (EASL 2020) proposed a new immune subtype based on the molecular characteristics of HCC, known as the “Immune-like” subtype. This novel subtype shares similarities with the immune active tumors, exhibiting strong interferon signaling activation and immune activity. However, it also presents CTNNB1 mutations. Specifically, interferon-gamma (IFN-γ) and its induced production of CXCL9 and CXCL10 contribute to the establishment of an inflammatory tumor microenvironment, which plays a role in the tumor’s response to ICIs therapy.Based on the aforementioned studies, an important review article published by Llovet et al. in 2022  providesIncreasing evidence suggests that achieving accurate subtyping of the tumor immune microenvironment in HCC requires attention to the spatial heterogeneity of the immune microenvironment. This spatial heterogeneity not only affects the accuracy of tumor immune microenvironment subtyping but also provides important clues for treatment decision-making .Research has found that small tumors in patients with multifocal HCC exhibit higher immune cell infiltration and immune pathway upregulation compared to larger tumors. This finding partially explains why small and large tumors within the same case may have different responses to anti-PD-1 therapy . Lancet Currently, the methods available for partially achieving phenotypic discrimination and spatial analysis of the HCC immune microenvironment include (I) in situ hybridization techniques, including fluorescence in situ hybridization (FISH) and multiplexed error-robust fluorescence in situ hybridization (MERFISH). The former enables qualitative or quantitative analysis of DNA, providing some rudimentary spatial information. The latter allows for highly multiplexed transcriptomic analysis at the single-cell level with spatial resolution. However, these methods can detect only a limited number of known target genes simultaneously . (II) SpAbove all, the classification of HCC tumor immune microenvironment helps to better characterize the distinct features of the immune microenvironment, thereby enabling more accurate prediction of tumor immunotherapy response. However, the current examination methods for immune cells, molecular phenotypes and spatial distribution have certain limitations, such as the following: (I) High technical requirements, limiting their implementation to specialized laboratories and making routine application in hospitals challenging. (II) High demands for tissue sample preservation and relatively complex processing steps during examination. (III) Limited coverage of the examined area, resulting in the loss of some in situ tissue structure and adjacent area information. (IV) Difficulty in obtaining gross pathological specimens from HCC patients receiving immune-related treatments, as results from needle biopsies can be influenced and may not fully reflect the spatial heterogeneity of the tumor. (V) Inability to achieve real-time dynamic monitoring of the tumor immune microenvironment in HCC patients.Therefore, there is an urgent need for an accurate, efficient, and convenient detection method to systematically and comprehensively evaluate the immune microenvironment phenotypes of individual tumors and their surrounding tissues. Especially, a useful tool is needed in clinical practice that would enable non-invasive prediction of the efficacy of immunotherapy and real-time dynamic monitoring of changes in the tumor immune microenvironment during treatment.In recent years, numerous researchers have made significant progress in the non-invasive evaluation of the immune microenvironment using magnetic resonance imaging (MRI), which has gradually become a research hotspot in this field. For instance, GPC-3, besides being a diagnostic biomarker for HCC, also plays a crucial role in the tumor immune microenvironment. It is considered as a potential target for the next generation of immune therapies . Chen etIn a study conducted by Professor Wang , Gd-EOB-max on fluorine-18 fluorodeoxyglucose PET-computed tomography (PET/CT) is associated with a poor clinical outcome and PD-L1 expression in patients with HCC. Furthermore, a one-armed antibody positron emission tomography tracer, 89ZED88082A, can visualize immunohistochemically confirmed CD8+ T cells in solid tumors by showing the tissue radioactivity localized areas [Positron emission tomography (PET) imaging has been proven to be a valuable noninvasive tool for evaluating the tumor microenvironment of HCC. For instance, Zhou et al.  reporteded areas .In addition to conventional imaging features for assessing the tumor immune microenvironment, radiomics has emerged as a viable approach in recent years for tumor diagnosis, microenvironment evaluation, and prognostic assessment . RadiomiIn the era of immunotherapy for HCC, the biggest challenge in clinical practice is the inability to predict which patients may have the greatest survival benefit from immunotherapy and which patients do not respond to immunotherapy. If we could accurately predict patients who are completely resistant to immunotherapy, it would not only save expensive medical resources, but more important, prevent these patients from receiving unnecessary immunotherapy drugs, thereby avoiding potential risks associated with drug side effects. Due to the immense tumor heterogeneity in HCC, tissue specimens obtained through pre-treatment biopsies are often unable to fully and accurately reflect the overall immune microenvironment of the tumor. Consequently, there is no current reliable biomarker in clinical practice to predict the efficacy of immunotherapy in HCC.As mentioned above, multi-parametric MR imaging contains a wealth of biological features, enabling non-invasive, comprehensive, and dynamic assessment of the immune microenvironment status in HCC . Kudo M.Presented studies have indicated that 20–30% of HCC patients categorized under the immune exclusion class may exhibit resistance to ICIs therapy. This resistance is attributed to diminished infiltration of CD8+ T-cells into tumors, caused by mutations in the WNT/β-catenin pathway and the absence of PD-L1 expression in cancer cells ,54. EvidRecently, multi-omics approaches have been used to elucidate the underlying mechanisms of immunotherapy response in HCC through the characterization of the tumor immune microenvironment features. Research has showcased the utilization of multi-omics signatures, specifically matrix metalloproteinase (MMP9), as a means to identify patients with distinct prognoses and varying responses to immunotherapy in HCC . This stOur analysis suggests that with the advancement of precision medicine, it is inadequate to simply employ radiomics and machine learning techniques for assessing the tumor immune microenvironment and predicting the efficacy of immunotherapy. Instead, it is crucial to explore stable radiomic features in these multicenter, large-scale, standardized HCC MRI datasets, and combine them with genomic, proteomic, and other pathological features to elucidate the biological mechanisms underlying these radiomic characteristics.Recently, artificial intelligence (AI) has emerged as a unique opportunity to improve the full spectrum of HCC clinical care, including HCC risk prediction, diagnosis, and prognostication . AdditioHowever, AI approaches, which commonly involve intricate layers of mathematical computations, pose significant challenges in gaining a comprehensive understanding of how data are transformed throughout the entire network. The pursuit of explainable AI is currently a growing area of research, with the primary objective of unveiling the inner workings of neural networks. Several research directions are being explored to demystify the ‘black boxes’ of neural networks. Key approaches include enhancing the transparency of networks, unraveling the semantics of different components, and generating post hoc explanations .Ensuring the interpretability of radiomics features and AI decisions is a shared concern across all current and future AI applications. Enabling various features of radiomics to have biological interpretability has always been a matter of great concern and a significant future research direction for radiologists. The notion of interpretability encompasses a specific array of techniques that empower users to grasp the inner workings and decision-making process of radiomics or AI models. It provides valuable insights into the essential factors influencing predictions and facilitates the understanding of potential biases. This transparency plays a crucial role in establishing the necessary trust to encourage medical professionals to embrace computer-aided devices that may be employed in the future .Multiple potential biomarkers for predicting ICI response of HCC have been explored and applied clinically. Yet there is no robust evidence to prove their clinical value in predicting immunotherapeutic response for patients with HCC. The identification of predictors which respond to ICIs is an urgent need and major challenge. Further studies could combine multi-parameter MRI with various AI techniques to develop and validate an effective tool for identifying the patients who would benefit from immunotherapy, and which can be used for predicting the tumor response to immunotherapy.


Cross-national research on cognitive aging inequality has largely concentrated on Western countries. It is unclear whether socioeconomic position (SEP) has similar effects on cognitive decline in emerging economies. We compared the association between life course SEP and cognitive function trajectories between China and England, the largest nation under state socialism and one of the oldest capitalist countries.n = 12,832) and the English Longitudinal Study of aging . Cognition z-scores were derived using comparable measures of memory and time orientation on 4 occasions. Life course SEP was self-reported by participants at baseline. Seven- to 8-year trajectories of cognition z-scores were estimated using latent growth curve modeling. Country- and gender-specific associations between childhood/adolescent deprivation, education, material wealth, and home ownership were evaluated in relation to model intercept (baseline level) and linear slope  of cognition.This cross-cohort study examined participants aged 50 years and older from the China Health and Retirement Longitudinal Study (z-score units per year); but compared with those with low education, Chinese men (b = 0.032) and women (b = 0.065) with high education had significantly slower declines than English men (b = −0.004) and women (b = 0.010) with high education.After multivariable adjustment, education was positively associated with the greatest differences in baseline cognition across country and gender. Education was further linked to a slower rate of cognitive decline (Despite substantial between-cohort differences in downstream and upstream determinants of dementia, education provided the greatest benefits to cognitive aging in England but particularly in China. Translational Significance: Given rapid worldwide population aging, a key policy aim is to target modifiable socioeconomic factors that can alleviate the burden of cognitive aging across countries with different levels of economic development and welfare systems. We compared cognitive trajectories and their association with four markers of socioeconomic position over the life course between China and England. This study found that education was most protective of cognitive function in both countries, suggesting that extending educational opportunities may promote cognitive health in older adults irrespective of the social context.Global dementia cases are anticipated to increase nearly threefold by 2050 due to rapid population aging, thus placing a new burden on the Global South as it grapples with the social and economic impacts of dementia in the coming decades . This raSocioeconomic position (SEP) over the life course refers to different aspects of social and economic conditions that inform an individual’s place in society from early life to old age . At partFirst, although individuals in disadvantaged SEPs are more likely to face worse circumstances and health in later life, one’s relative social standing is rooted in the broader structural context . CompariSecond, the cumulative effects of life course SEP on cognitive aging may be starker in some contexts. Early life deprivation lowers childhood cognitive ability, creating deficits that track into adulthood . This asA disadvantaged SEP over the life course has been associated with worse cognitive function and faster decline . With grWe analyzed nationally representative samples from two sister studies of aging, the China Health and Retirement Longitudinal Study (CHARLS: ≥45 years)  and the CHARLS and ELSA had three identical cognitive tests across waves. Participants recalled as many words as possible immediately after a 10-word list was read  and after a short delay . The number of correctly remembered words from both trials was tallied to a score from 0 to 20. Participants also reported the: (i) day of month; (ii) month; (iii) year; and (iv) day of the week. Accurate responses were summed to a score from 0 to 4. The sum of the three cognitive tests constituted the main study outcome measure (0–24). Anterograde memory (referring to the ability to learn and retain newly encountered information) and orientation in time are fundamental cognitive functions  that decAdditional tests included in the cognitive examinations were not identical between CHARLS and ELSA. Yet, these cognitive tests encompassed other important domains, such as attention  and visuoconstruction (pentagon copy task = 0–3) in CHARLS and language in ELSA  were used to calculate z-scores on four occasions for cognition  and total cognition . This is a widely implemented approach in longitudinal analyses of cognition z-scores ; body mass index (BMI) categories ; smoking status ; and alcohol drinking frequency in the past year . Health covariates comprised the number of limitations in up to 5 activities of daily living ; self-rated hearing; probable depression using thresholds for the 10-item Center for Epidemiological Depression (CES-D) scale  scale  in CHARLD) scale  in ELSA D) scale . Althougz-scores across four occasions estimated the baseline level (intercept) and the rate of change (slope) for every year of follow-up. Longitudinal changes in cognition were slightly curvilinear in both studies, therefore, trajectories were modeled with a linear and quadratic slope.Prospective cognition trajectories  were assessed using latent growth curve modeling (LGCM) in Mplus (version 7.4). Linear regression of the cognition p values for interactions between gender with education and wealth, respectively, on the intercept (<.0001) and linear slope  in CHARLS, the comparative analyses were gender stratified.Sequential study-specific models estimated the intercept and linear slope with the following covariates: individual SEP measure, age, and age squared ; Model 1 covariates plus marital status, BMI, smoking status, and drinking frequency (Model 1 + lifestyle factors); Model 1 covariates plus limitations in ADLs, hearing, probable depression, cardiovascular disease, hypertension, and diabetes ; and all covariates (Model 2). Although the quadratic slope was statistically significant, it was not regressed on covariates due to its small effect size and little inter-individual variability. Owing to Cognition scores were completed on four occasions for a portion of the main analytic samples . Longitudinal participation patterns and associated mean scores suggested that individuals with better cognition were somewhat more likely to be followed up in both studies . To addrz-scores, which comprised all cognitive functions available over the study period in CHARLS (plus attention and visuoconstruction) and ELSA (plus language).The main analysis is based on identical measures of anterograde memory and time orientation. To assess whether the main findings are sensitive to other cognitive domains, we replicated the main LGCMs using total cognition At baseline, English adults had higher memory and time orientation scores than those in China , resultiz-scores were 0.037 units  lower among Chinese women who reported deprivation and were partially explained by health conditions (Model 2B) rather than lifestyle factors (Model 2A). Weaker associations in England may result from inadequate power because 3% of ELSA participants reported severe financial hardship before adulthood. Although deprivation was more widespread in China, these circumstances were not associated with faster age-related cognitive declines. Cognitive decline rates per year were similarly weak by deprivation in England.Individuals who experienced deprivation in childhood/adolescence had lower cognition at baseline, but age-adjusted associations (Model 1) were strong only in Chinese women  and 3. Cz-score units per year) associated with education remained robust after full adjustment in Chinese men  and women . Education was less protective against cognitive aging in England as the linear decline in z-score units per year was more marginal, and only significantly slower for English women with high education before  and after full adjustment .Education most strongly predicted baseline cognition across context and gender. Compared with wealth and home ownership, effect sizes by education were largest for both genders in both countries before and after adjustment. Baseline inequalities were partially explained by lifestyle and related health factors, although lifestyle played a greater role in England. Higher educational attainment was associated with slower cognitive declines in China, particularly among women. The slower rate of cognitive decline .Home ownership was associated with cognition in England, but not in China. English adults who did own their current homes had worse baseline cognition, partly due to lifestyle and health factors. Although non-home ownership was also associated with faster cognitive declines in English adults, these prospective effects were largely attenuated after further adjustment.As education was the strongest predictor of cognition, z-scores were largely similar to the main results, indicating that the differential effects of SEP are impervious to cognitive functions which were excluded from the main analysis , and wealth (England) were more strongly associated with baseline cognition than rates of decline . Contrard to men . Diverged to men  which wad to men .The Global South has scarcely featured in cross-national prospective studies of cognitive aging inequalities. Although recent international studies, including China, reported that cross-sectional associations of cognitive function appeared stronger with education than with income , this doAlthough we did not set out to examine gender differences, wider educational inequalities in cognitive aging observed among older Chinese versus English women draw our attention to gender inequalities pertaining to education and other life chances. As CHARLS women had much lower levels of education than CHARLS men, as well as ELSA women, this may explain why Chinese women, particularly those with the lowest education, had the fastest declines in cognitive function. Cross-national literature on gender and health point to context-specific gendered vulnerabilities pertaining to social roles and social positions within different spheres of domestic, social, and occupational life. Female roles are often associated with certain limitations which can lead to disadvantaged social positions, although this differs from society to society . BloombeThe salutary role of education in cognitive aging brings to mind global secular improvements in formal schooling. Although the broader literature has linked the decrease in dementia incidence across consecutive cohorts with population improvements in education, health care, nutrition and other lifestyle changes, a reversal of these cohort trends is plausible given increasing obesity, diabetes, and social isolation, coupled with declining levels of physical activity . The anaLast, our findings indicated that lifestyle factors such as BMI, smoking, and drinking behaviors had less explanatory power between life course SEP and cognitive aging in China than in England. These findings illustrate East–West differences in the social patterning of health behaviors which are largely driven by cultural and social norms . Althougz-score, which meets the assumption that being 1 SD below the mean in CHARLS is identical to being 1 SD below the mean in ELSA. Absolute cognitive levels were directly comparable between countries, thereby facilitating the comparison of within-country associations with SEP. Although the main findings may not fully reflect cognitive aging inequalities in each setting given the multidimensional nature of the cognitive function, sensitivity analysis confirmed that within-country associations with SEP did not materially change when using the total cognition z-score  available in CHARLS and ELSA.Analogous data on anterograde memory and time orientation were combined to create a harmonized cognition Data on adult SEP were self-reported by participants at baseline. Material wealth and home ownership in later life indicate economic resources accrued over the life course and are important markers of older people’s SEP because social advantage amasses over time . EducatiAttrition bias may have underestimated cognitive inequality, but all participants with at least one cognition score over the follow-up were retained using FIML. Although a Danish study found that the association between higher education and slower cognitive decline over 10 years was not influenced by selective dropout due to death , the comNotwithstanding considerable differences in underlying health status and social context, chronic disease antecedents, and sociodemographic patterns of health behaviors , educatiigad064_suppl_Supplementary_MaterialClick here for additional data file.