Early Development Research Articles

#1214 Post-kidney transplant cancer: incidence, risk factors and future perspectives. Real-world analysis of a single Italian center

Abstract Background Our retrospective observational study aims to describe the epidemiology of cancer after KTx and to investigate its risk factors and the impact on therapeutic management and survival, in a big monocentric cohort of kidney transplant patients (KT-ps). The association between a specific modification of immunosuppressive therapy after cancer diagnosis and survival outcomes will be analyzed. Methods Retrospective collection of data in KTx-ps transplanted between January 2004 and December 2021, subjected to outpatient follow-up with a median follow-up of 7 [1-19] years. Results We included 930 KTx-ps with mean age at KTx of 49 years. 91% had received pre-TR dialysis, for an average of 52 months. The majority had received KTx from a deceased donor (84%). 74% had undergone induction therapy with Basiliximab and 26% ATG. Maintenance therapy included in most cases the use of steroids (87%), calcineurin inhibitors (tacrolimus 92%, ciclosporin 8%), and mycophenolate (94.6%). Patients with at least one cancer (CAN+) were 177 (19%). The mean time from TR to oncological diagnosis was 83 months. Non-melanoma skin cancers (NMSC) were the most common tumors (55%), while solid tumors were observed in 38.6% of cases. Deceased donor was more represented in CAN+. CAN+ were older and with higher BMI. They had greater presence of vasculitis as basic nephropathy. In the period between 2016 and 2021, induction therapy with ATG was significantly associated with CAN+. In multivariable analysis, ATG emerged as an independent risk factor for CAN+ (p=0.014) and, in survival analyses, ATG was strongly and significantly related to the earlier development of CAN+ (p=0.010). During follow up, cancer-related causes were the second cause of mortality (23%); the average time from cancer diagnosis to death was 23 months. The median survival from TR was 148 (CAN+) and 82 months (CAN−). After oncological diagnosis, significant evidence on tumor survival was derived from the shift to mTOR inhibitors compared to the group with definitive drug suspension (p=0.004). Conclusion The data reported by our study confirm the relevance of cancer as a complication in KTx. ATG represent an independent risk factor for cancer; in this context, a personalized choice of immunosuppressive therapy to be proposed at the time of TR represents a crucial point in the prevention of neoplastic risk. In the KTx-ps, therapeutic strategies with antineoplastic agents and management of immunosuppression therapy after malignancy remain important research perspectives in the near future.

Cerebrovascular miRNAs Track Early Development of Alzheimer's Disease and Target Molecular Markers of Angiogenesis and Blood Flow Regulation.

Alzheimer's disease (AD) is associated with impaired cerebral circulation which underscores diminished delivery of blood oxygen and nutrients to and throughout the brain. In the 3xTg-AD mouse model, we have recently found that > 10 cerebrovascular miRNAs pertaining to vascular permeability, angiogenesis, and inflammation (e.g., let-7d, miR-99a, miR-132, miR-133a, miR-151-5p, and miR-181a) track early development of AD. Further, endothelial-specific miRNAs (miR-126-3p, miR-23a/b, miR-27a) alter with onset of overall AD pathology relative to stability of smooth muscle/pericyte-specific miRNAs (miR-143, miR-145). We tested the hypothesis that cerebrovascular miRNAs indicating AD pathology share mRNA targets that regulate key endothelial cell functions such as angiogenesis, vascular permeability, and blood flow regulation. As detected by NanoString nCounter miRNA Expression panel for 3xTg-AD mice, 61 cerebrovascular miRNAs and respective mRNA targets were examined using Ingenuity Pathway Analysis for canonical Cardiovascular (Cardio) and Nervous System (Neuro) Signaling. The number of targets regulated per miRNA were 21±2 and 33±3 for the Cardio and Neuro pathways respectively, whereby 14±2 targets overlap among pathways. Endothelial miRNAs primarily target members of the PDE, PDGF, SMAD, and VEGF families. Individual candidates regulated by≥4 miRNAs that best mark AD pathology presence in 3xTg-AD mice include CFL2, GRIN2B, PDGFB, SLC6A1, SMAD3, SYT3, and TNFRSF11B. miRNAs selective for regulation of endothelial function and respective downstream mRNA targets support a molecular basis for dysregulated cerebral blood flow regulation coupled with enhanced cell growth, proliferation, and inflammation.

Insights from applications of the RSTM tool for coupled CFD-activation fluid simulation

Accurate prediction of the activation of fluids flowing under irradiation is important for the timely development of fusion technology. One of the major current issues is the cooling water of ITER, which becomes activated by plasma neutrons during nuclear operations, thus raising a number of radiological implications such as radiation effects on sensitive equipment, compliance with radiological protection zoning, and compliance with radioactivity limits in regulations of pressure equipment and effluents. Further significant applications are the activation of other service fluids in ITER and of the LiPb in both ITER and DEMO breeding blanket modules.To avoid systematic uncertainties, simulation of the activation of fluids flowing in arbitrarily complex 3D geometry, flow regimes and neutron fields requires full coupling of activation with fluid-dynamics physics models. No such tools were available until recently: the Radio-Species Transport Model (RSTM), based on the well-established ANSYS Fluent® UDS methodology, is one conceived and developed at F4E. Here we review the methodology and capabilities of the RSTM, as well as earlier development, validation, benchmarking and application activities. We then report currently ongoing further applications and benchmarking being performed in collaboration with specific tasks of the EUROFusion programme Preparation for ITER Operation. Computations, results and comparison with other methodologies for several cases of interest are presented and discussed.

Systemic lupus erythematosus with juvenile onset: current status of the problem (literature review)

Patients with juvenile-onset systemic lupus erythematosus (jSLE) account for up to 25% of all SLE patients. The main difference between jSLE and SLE in adults is the greater role of genetic factors in the pathogenesis, higher activity, earlier development of complications and the need for more aggressive immunosuppressive therapy, which allows us to consider the onset of the disease in childhood as a special phenotype of SLE. The relevance of the study of jSLE arises from the variability of clinical manifestations and the unpredictability of the course, the difficulty of early diagnosis, the rapid development of organ damage and the unfavorable life prognosis.The article presents the most important modern data on the diagnosis, classification, features of the clinical picture and treatment approaches of jSLE from a practical point of view.

Power Generation Enhancement through Latching Control for a Sliding Magnet-Based Wave Energy Converter

A Surface-Riding Wave Energy Converter (SR-WEC) featuring a sliding magnet inside a pitching cylindrical hull is investigated as an easily deployable small power device to support small-scale marine operations. This study extends the earlier development of the system by authors to enhance power performance through the application of end spring and latching control. The inclusion of springs at the tube’s end enhances the magnet release and travel speeds as well as the average power output compared to systems without them. Further improvement of power output can also be achieved by employing optimal latching control. We introduced constant-angle and variable-angle unlatching strategies to determine optimal parameters in combination with passive and reactive power take-off (PTO) controls to assess their effectiveness. The optimized latching control and end spring can increase 60–80% more power output compared with the case without them under certain PTO damping. Additionally, we discussed the effects of limiting peak powers and associated energy leaks with latching.

Investing in socio-emotional skills during early childhood to achieve capabilities and fight poverty: overcoming socioeconomic vulnerabilities in Brazil through Criança Feliz

This paper aims to discuss early childhood social-emotional development as a means to achieve capabilities and reduce poverty and inequality. Social-emotional skills provide the necessary cognitive tools to support individuals during choices and decisions that will affect socioeconomic status. Therefore, the connection between the socio-emotional skills approach and the capabilities must be established. When developed and mastered, the social-emotional skills improve opportunities and promote new capabilities to achieve desirable results (capabilities to function). If this development starts during early childhood, it ensures the right to exercise agency power, improving future economic results. For that reason, developing these skills in disadvantaged populations is a necessary condition to promote freedom and equality of opportunity, ensuring inclusive socio-economic development. The goals of "Programa Criança Feliz" (PCF), an early childhood social-emotional development program for disadvantaged children launched in October 2016 in Brazil, are analyzed in this paper through the connection between socio-emotional skills and the capabilities approach. This program integrates public policies that reduce the weakness of will and provide skills to overcome socioeconomic vulnerabilities in Brazil. Therefore, this paper contributes through the analysis of PCF, and the connection of the development of socio-emotional skills during early childhood to achieve capabilities and fight poverty.

Examining timing effects in the intergenerational transmission of anxiety and depressive symptoms: A genetically informed study.

The present study examined genetic, prenatal, and postnatal environmental pathways in the intergenerational transmission of anxiety and depressive symptoms from parents to early adolescents (when these symptoms start to increase), while considering timing effects of exposure to parent anxiety and depressive symptoms postnatally. The sample was from the Early Growth and Development Study, including 561 adopted children (57% male, 55% White, 13% Black/African American, 11% Hispanic/Latine, 20% multiracial, 1% other; 407 provided data in early adolescence) and their birth (BP) and adoptive parents (AP). Using a trait-state-occasion model with eight assessments from child ages 9 months to 11 years, we partitioned trait-like AP anxiety and depressive symptoms from time-specific fluctuations of AP anxiety and depressive symptoms. Offspring anxiety and depressive symptoms were assessed at 11 years (while controlling for similar symptoms at 4.5 years). Results suggested that time-specific fluctuations of AP1 (mostly mothers) anxiety/depressive symptoms in infancy (9 months) were indirectly associated with offspring anxiety/depressive symptoms at 11 years via offspring anxiety/depressive symptoms at 4.5 years; time-specific fluctuations of AP1 anxiety/depressive symptoms at child age 11 years were concurrently associated with offspring anxiety/depressive symptoms at 11 years. AP2 (mostly fathers) anxiety/depressive symptoms were not associated with offspring symptoms. Genetic and prenatal influences measured by BP internalizing problems were not associated with offspring symptoms. Results suggested infancy and early adolescence as developmental periods when children are susceptible to influences of parent anxiety and depressive symptoms. Preventive interventions should consider time-specific fluctuations in parent anxiety and depressive symptoms during these developmental periods. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

A maturational frequency discrimination deficit may explain developmental language disorder.

Auditory perceptual deficits are widely observed among children with developmental language disorder (DLD). Yet, the nature of these deficits and the extent to which they explain speech and language problems remain controversial. In this study, we hypothesize that disruption to the maturation of the basilar membrane may impede the optimization of the auditory pathway from brainstem to cortex, curtailing high-resolution frequency sensitivity and the efficient spectral decomposition and encoding of natural speech. A series of computational simulations involving deep convolutional neural networks that were trained to encode, recognize, and retrieve naturalistic speech are presented to demonstrate the strength of this account. These neural networks were built on top of biologically truthful inner ear models developed to model human cochlea function, which-in the key innovation of the present study-were scheduled to mature at different rates over time. Delaying cochlea maturation qualitatively replicated the linguistic behavior and neurophysiology of individuals with language learning difficulties in a number of ways, resulting in (a) delayed language acquisition profiles, (b) lower spoken word recognition accuracy, (c) word finding and retrieval difficulties, (d) "fuzzy" and intersecting speech encodings and signatures of immature neural optimization, and (e) emergent working memory and attentional deficits. These simulations illustrate many negative cascading effects that a primary maturational frequency discrimination deficit may have on early language development and generate precise and testable hypotheses for future research into the nature and cost of auditory processing deficits in children with language learning difficulties. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Reciprocal Effects Between Negative Parenting and Children's Callous-Unemotional Traits From Mid to Late Childhood.

The role of negative parenting in the development of callous-unemotional (CU) traits remains unclear. Both negative parenting and CU traits are influenced by genetic and environmental factors. The authors used genetically informed longitudinal cross-lagged models to examine the extent to which reciprocal effects between negative parenting and children's CU traits in mid-to-late childhood are genetic versus environmental in origin. In 9,260 twin pairs from the Twins Early Development Study, the authors estimated cross-lagged effects between negative parenting (discipline and feelings) and children's CU traits in mid (ages 7-9) and late (ages 9-12) childhood. CU traits were strongly heritable and stable. Stability was explained largely by genetic factors. The influence of negative parenting on the development of CU traits was small and driven mostly by genetic and shared environmental factors. In mid childhood, the influence of children's CU traits on subsequent negative parenting (i.e., evoked by children's CU traits) was also small and mostly genetic in origin. In late childhood, CU traits showed no effects on negative parental discipline and small effects on negative parental feelings, which reflected mostly shared environmental factors. In mid-to-late childhood, genetic factors strongly influenced the development of CU traits, whereas environmental effects of negative parenting were small. Negative parenting was also relatively unaffected by CU traits. The small reciprocal effects originated mostly from genetic and shared environmental factors. Therefore, repeated intensive interventions addressing multiple risk factors rather than negative parenting alone may be best positioned to support families of children with CU traits across development.

Nutritional dynamics in early development of Asian Openbill: A study of hatchling and nestling feeding patterns

Nutrition plays a crucial role in avian development, growth, and health. Inadequate nutrition can have an impact on immune systems and increase susceptibility to diseases. However, there is a paucity of studies that quantify the nutrition provided to young birds. We studied the nutrients delivered to six nests, especially to the hatchling and nestling stages of the Asian Openbill (Anastomus oscitans), using CCTV cameras and spotting scope in Eastern Nepal. Simultaneously, we collected the major food of the Asian Openbill, specifically Filopaludina and Pila spp., and assessed their nutrient content value to estimate the amount of nutrients delivered to the young birds. Asian Openbills feed Filopaludina spp. to hatchlings and both Filopaludina and Pila species to nestlings across all nests. The amount of food supplied to each nest during the nestling stage (272.4 to 386.4 g/day) was approximately three times higher than that during the hatchling stage (87.7 to 128.01 g/day). Energy was primarily derived from protein, followed by fat, and carbohydrate, with variations observed between both nesting stages. Our study provides baseline data on nutrient delivery patterns during two nesting stages of the Asian Openbill in Eastern Nepal, highlighting their adaptability and flexibility in meeting nutritional requirements.

Oxygen levels affect oviduct epithelium functions in air–liquid interface culture

Key reproductive events such as fertilization and early embryonic development occur in the lumen of the oviduct. Since investigating these processes in vivo is both technically challenging and ethically sensitive, cell culture models have been established to reproduce the oviductal microenvironment. Compartmentalized culture systems, particularly air–liquid interface cultures (ALI; cells access the culture medium only from the basolateral cell side), result in highly differentiated oviduct epithelial cell cultures. The oxygen (O2) tension within the oviduct is 4–10% across species, and its reduced O2 content is presumed to be important for early reproductive processes. However, cell culture models of the oviduct are typically cultivated without O2 regulation and therefore at about 18% O2. To investigate the impact of O2 levels on oviduct epithelium functions in vitro, we cultured porcine oviduct epithelial cells (POEC) at the ALI using both physiological (5%) and supraphysiological (18%) O2 levels and two different media regimes. Epithelium architecture, barrier function, secretion of oviduct fluid surrogate (OFS), and marker gene expression were comparatively assessed. Under all culture conditions, ALI-POEC formed polarized, ciliated monolayers with appropriate barrier function. Exposure to 18% O2 accelerated epithelial differentiation and significantly increased the apical OFS volume and total protein content. Expression of oviduct genes and the abundance of OVGP1 (oviduct-specific glycoprotein 1) in the OFS were influenced by both O2 tension and medium choice. In conclusion, oviduct epithelial cells can adapt to a supraphysiological O2 environment. This adaptation, however, may alter their capability to replicate in vivo tissue characteristics.

Весеннее развитие черемухи обыкновенной как индикатор временных состояний ландшафтов (Свердловская область)

В статье представлены результаты исследования весеннего развития черемухи обыкновенной при проведении Единого фенологического дня 15 мая на территории Свердловской области за 2012–2023 гг. Изучение сезонного состояния вида при помощи описательного метода позволило определить географические закономерности наступления фенологических фаз и скорости продвижения явления в пространстве в пределах природных зон и подзон в зависимости от факторов окружающей среды. Выявлено более раннее развитие черемухи на юго-востоке и юго-западе области, далее фронт явления продвигается на север и северо-запад. Различия между северными и южными районами территории, в среднем, составляют 6–7 баллов. В результате наблюдений получена характеристика сезонного развития объекта, индицирующего состояние природных комплексов в краткой математической форме. Максимальное опережение развития выявлено в лесостепной зоне Русской равнины и Западной Сибири в границах исследуемого региона. В горных районах области явления наступают с заметным отставанием в 6 баллов по сравнению с равнинными. Скорость прохождения явления в разных под зонах колеблется от 0,8 до 3 баллов/100 км. Опыт такого рода многолетних исследований для других районов страны может помочь выявить тенденции влияния изменений климата на адаптационные механизмы отдельных видов растений-феноиндикаторов.

A retrospective cohort study analyzing the changes in early childhood development during the COVID-19 pandemic

ObjectiveTo investigate early childhood development (ECD) outcomes in different subgroups before and during the COVID-19 pandemic. Study designA retrospective cohort study of children 3–58 months of age whose caregivers completed a Survey of Well-being of Young Children (SWYC) as part of a well child visit (WCC). The data were divided into two phases: pre-pandemic (September 2018 – February 2020), and during pandemic (September 2020 – February 2022). The difference in the proportion of forms with Meets Expectations interpreted scores on the SWYC Developmental Milestones pre-pandemic versus during the pandemic timeframe overall and among subgroups were reported. Hypotheses were tested using logistic regression with repeated measures. Results14,550 patients were included in the sample for analysis with 52,558 SWYC form observations. There was no difference in the odds of a Meets Expectations interpreted score before and after the pandemic for the entire sample, OR 0.99 (95 % CI: 0.94–1.04). There was evidence of decreased odds of an interpreted score of Meets Expectations for the following subgroups: male, Hispanic/Latino ethnicity, ages of 24, 30 or 36 months at WCC, Medicaid insurance, 2nd HOUSES Quartile, requiring interpreter, single parent household, young maternal age, maternal substance abuse, and race identified as Native Hawaiian/Pacific Islander, American Indian/Native Alaskan or Other. ConclusionDecreased odds of meeting developmental milestones during the pandemic were evident in certain high risk sub-groups revealing unequal distribution of suboptimal developmental outcomes within our population during the pandemic that may be exacerbating existing inequities impacting development in children.

Early development of infants born prematurely who were conceived through assisted reproductive technologies

ObjectiveThis study aims to determine physical and psychomotor development characteristics during the first 24 months of life in prematurely born children conceived with assistive reproductive technologies and to determine specific risk factors for deviation from typical development. Patients and methodsThe study included 308 prematurely born children (154 conceived with assistive reproductive technologies – ART group and 154 conceived naturally – NC group) hospitalized at the Institute for the Health Care of Children and Adolescents of Vojvodina in Serbia. The physical and psychomotor development was monitored during the first two years of life. ResultsBirth weight was statistically significantly higher in the NC group than in the ART group, and IUGR was more often present in the ART group than in the NC group of newborns. The mean values of body weight, length, and head circumference of children from both groups did not differ statistically significantly at 18 months. At 12 and 18 months, children from the ART group significantly more often had below-average scores on the test for assessment of psychomotor development (corrected GDQ <90) than children from the NC group, but not at 24 months. Logistic regression analysis has shown that at 12 and 18 months, multiple pregnancy is an independent risk factor for lower GDQ. ConclusionAt the age of 12 months, prematurely born children conceived with ART do not differ in physical development from prematurely born children after natural conception. At 12 and 18 months, prematurely born children conceived with ART achieve lower global developmental scores than children conceived naturally, but not at 24 months of age. Multiple pregnancies are an independent risk factor for lower achievement on the test for assessment of psychomotor development in prematurely born children.

Better in sync: Temporal dynamics explain multisensory word-action-object learning in early development.

We investigated the temporal impact of multisensory settings on children's learning of word-object and action-object associations at 1- and 2-years of age. Specifically, we examined whether the temporal alignment of words and actions influenced the acquisition of novel word-action-object associations. We used a preferential looking and violation of expectation task in which infants and young children were first presented with two distinct word-object and action-object pairings either in a synchronous (overlapping in time) or sequential manner (one after the other). Findings revealed that 2-year-olds recognized both, action-object and word-object associations when they first saw the word-action-object combinations synchronously, but not sequentially, as evidenced by looking behavior. 1-year-olds did not show evidence for recognition for either of the word-object and action-object pairs, regardless of the initial temporal alignment of these cues. To control for individual differences, we explored factors that might influence associative learning based on parental reports of 1- and 2-year-olds development, however, developmental measures did not explain word-action-object associative learning in either group. We discuss that while young children may benefit from the temporal alignment of multisensory cues as it enables them to actively engage with the multisensory content in real-time, infants may have been overwhelmed by the complexity of this input.

Abstract 7003: Microfluidic platform for DNA methylation profiling towards early detection of ovarian cancer

Abstract We present a microfluidic platform for molecule-by-molecule detection of heterogeneous epigenetic patterns of rare tumor-derived DNA by highly parallelized digital melt. The platform digitizes DNA molecules in a 4-module, 40,000 1-nL array microfluidic device and observes sequence-dependent fluorescence changes during temperature ramping. The platform is applied to quantify DNA methylation heterogeneity of a biomarker panel within Pap specimens in a small cohort of healthy and ovarian cancer patients, and demonstrates an accuracy of 0.9. DNA hypermethylation of tumor-suppressor genes is an epigenetic phenomena that occurs early in tumorigenesis. Variable methylation patterns are observed in ovarian cancer precursor tissue, and trace amounts of ovarian-tumor-derived DNA can be found in Pap specimens, albeit at low frequencies (&amp;lt;0.01%). This suggests that methylation profiles of DNA extracted from routinely-obtained Pap specimens could contain early cancer biomarkers. However, due to the scarcity of these molecules, current techniques, such as sequencing and digital PCR (dPCR), have technical limitations precluding their ability to profile methylation patterns reliably. Sequencing, although comprehensive, has limited sensitivity, undermining its utility for routine detection of fractions below 0.1%. dPCR achieves high sensitivity, but is limited to known sequences, thus cannot provide comprehensive analysis of molecular heterogeneity. To overcome these limitations, we developed a technique called digital high resolution melt (dHRM), a thermodynamics-based approach to detecting molecular variability by observing the sequence-specific release of a DNA-intercalating dye under a thermal ramp. dHRM can discriminate methylation patterns on CpG-by-CpG basis and detect methylated epialleles at frequencies as low as 0.00005%. Here, DNA from Pap specimens was loaded on a microfluidic chip that digitizes rare target molecules into individual reaction chambers. A thermal-optical platform was developed to perform parallelized dPCR and dHRM. Methylation patterns of a biomarker panel were analyzed to produce a probability score that the Pap specimen contained tumor-derived DNA. 12 healthy and 12 ovarian cancer Pap specimens were assessed for methylation heterogeneity of 9 genes. Optimal methylation density thresholds were determined, and data above each threshold was combined into a single score for each possible combination of biomarkers. In this small cohort, methylation heterogeneity analysis of just 2 loci produced an area under the receiver-operator characteristics curve (AUC) of 0.9. This highly sensitive methylation profiling technology demonstrates promising utility towards early cancer detection. Future work aims to expand the testing cohort for ovarian cancer detection and permit further studies on the impact of methylation heterogeneity on early cancer development. Citation Format: Chrissy O'Hersey, Yang Zhao, Thomas R. Pisanic, Tian-Li Wang, Ie-Ming Shih, Tza-Huei Jeff Wang. Microfluidic platform for DNA methylation profiling towards early detection of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7003.

Abstract 5437: Chemoproteomics-enabled discovery of covalent inhibitors targeted at AGPAT4: Unravelling tumor lineage plasticity to overcome drug resistance in hepatocellular carcinoma

Abstract The liver is a unique organ that is responsible for many metabolic functions. During hepatocellular carcinoma (HCC) development, these metabolic machineries are extensively reprogrammed to support the insatiable nutrient requirements of HCC. Tumor lineage plasticity, a recognized hallmark of cancer, is a phenomenon in which tumor cells co-opt developmental pathways to attain cellular plasticity, enabling them to evade targeted therapeutic interventions. Cancer cells can reprogram their metabolic pathways to match their increased metabolic needs for cancer cell survival under adverse conditions. Identifying novel metabolic targets related to stemness can offer promising strategies for targeting cancer stemness roots and hence to kill and control their growth. Through pathway enrichment analysis of genes that linked metabolism and stemness, we identified an aberrant glycerophospholipid metabolism signature, with AGPAT4 ranking as the top-hit. AGPAT4 upregulation in HCC is strongly correlated with aggressive clinical features, including survival, metastasis, and stemness signatures. AGPAT4 expression peaked during early liver progenitor development, decreased during hepatocyte maturation and progressively increased from well-differentiated to poorly differentiated HCCs. Enrichment of AGPAT4 in HCC is mediated by promoter binding of SOX9 to drive AGPAT4 transcriptional activity. AGPAT4 inhibition can mitigate tumor initiation, self-renewal, metastasis, and sorafenib resistance. Mechanistic studies revealed an AGPAT4-mediated phosphatidic acid production axis that promotes HCC through the regulation of mTOR signaling. Inhibition of Agpat4 by AAV8 shRNA reduced tumorigenicity and stemness, and sensitized HCC tumors to sorafenib. AGPAT4 overexpression can predict sorafenib response in clinical settings. Through chemoproteomics screening of a cysteine-reacting compound library using activity-based protein profiling, a cysteine-reacting compound with high binding affinity and selectivity towards AGPAT4 was identified and found to work synergistically with sorafenib to suppress HCC, as demonstrated in HCC patient-derived tumor xenograft (PDTX) models. Toxicity analysis through histological examination of organs, body weight measurements, and biochemical tests revealed minimal toxicity associated with the covalent inhibitor. In conclusion, AGPAT4 is a novel metabolic driver of oncogenic stemness, dedifferentiation, and metastasis in HCC. AGPAT4-induced tumor lineage plasticity may represent an Achilles heel for HCC treatment, and inhibition of AGPAT4 may widen the therapeutic window for sorafenib treatment in the clinic. Citation Format: Kai-Yu Ng, Tin-Yan Koo, Tsz-Lok Fong, Ya Gao, Tin-Lok Wong, Yuan Gao, Jing-Ping Yun, Xin-Yuan Guan, Ming Liu, Clive YS Chung, Stephanie Ma. Chemoproteomics-enabled discovery of covalent inhibitors targeted at AGPAT4: Unravelling tumor lineage plasticity to overcome drug resistance in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5437.

Reversible Influence of Hemipiperazine Photochromism on the Early Development of Zebrafish Embryo.

This study explores the potential of controlling organismal development with light by using reversible photomodulation of activity in bioactive compounds. Specifically, our research focuses on plinabulin 1, an inhibitor of tubulin dynamics that contains a photochromic motif called hemipiperazine. The two isomeric forms, Z-1 and E-1, can partially interconvert with light, yet show remarkable thermal stability in darkness. The Z-isomer exhibits higher cytotoxicity due to stronger binding to α-tubulin's colchicine site. The less toxic E-1 form, considered a "pro-drug", can be isolated in vitro and stored. Upon activation by blue or cyan light, it predominantly generates the more toxic Z-1 form. Here we demonstrate that 1 can effectively photomodulate epiboly, a critical microtubule-dependent cell movement during gastrulation in zebrafish embryos. This research highlights the potential of photomodulation for precise and reversible control of cellular activities and organismal development.

Abstract 2413: Mutated TP53 prevalence in EGFR-mutated advanced non-small cell lung cancer patients with brain metastases

Abstract Introduction: Approximately 50% of patients with EGFR-mutated advanced non-small cell lung cancer (NSCLC) develops brain metastases (BM), which is associated with a poor prognosis. Mutations in TP53 are associated with earlier development of resistance to EGFR tyrosine kinase inhibitors, but it is unclear whether patients who develops BM have a higher prevalence of TP53 mutations. Analyses of circulating tumor DNA (ctDNA) in blood have been established as a good alternative to tissue biopsies to assess the genomic landscape of NSCLC. In this study, we aim to analyze ctDNA from patients with advanced EGFR-mutated NSCLC to investigate the prevalence of TP53 mutations in patients with BM, and to explore whether patients with BM exhibit a distinct ctDNA profile compared to patients without. Materials and Methods: Plasma samples collected before treatment commenced from 97 of the 100 patients with EGFR-mutated advanced NSCLC enrolled in the First-line Treatment With Osimertinib in EGFR-mutated Non-small Cell Lung Cancer study (The FIOL study: NCT03804580) were analyzed. Patients were split into cohorts with (n=44) and without BM (n=53) at baseline. The cell-free DNA was isolated and sequenced with targeted next-generation sequencing with the AVENIO ctDNA Surveillance Panel (Roche) containing 197 lung cancer-related genes. Results: Mutations in ctDNA were found in 83 patients (85.6%), with 6 patients with BM having no ctDNA mutations and 8 patients without BM having no ctDNA mutations. Besides EGFR mutations, TP53 was the most frequently mutated gene, with 42 patients (43.3%) harboring TP53 mutations. There was no significant difference in the prevalence of TP53 mutations between the cohort with BM (n=23) and the cohort without (n=19), (p=0.15). There was no difference in the number of identified mutations in the two cohorts (BM: median: 2 (range: 0-6), without BM: median: 2 (range: 0-11), p=0.71). Patients with BM had a numerically lower ctDNA level (median: 45.5 mutant molecules/mL), than patients without BM (median: 75.9 mutant molecules/mL), though the difference was not statistically significant (p=0.68). Conclusion: Prevalence of TP53 mutations in plasma collected before osimertinib treatment initiation in advanced EGFR-mutated NSCLC was similar between patients with and without BM at baseline. Furthermore, there was no difference in the ctDNA profile between these two cohorts. Future experiments will clarify the impact of TP53 mutations in patients with or without BM. Citation Format: Simone Stensgaard, Inger Johanne Z. Eide, Elin Marie Stensland, Åslaug Helland, Simon Ekman, Karin H. Hansen, Saulius Cicenas, Bjørn Henning Grønberg, Peter Meldgaard, Boe S. Sørensen, Odd Terje Brustugun. Mutated TP53 prevalence in EGFR-mutated advanced non-small cell lung cancer patients with brain metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2413.

Postnatal survival of phrenic motor neurons is promoted by BDNF/TrkB.FL signaling.

The number of motor neurons (MNs) declines precipitously during the final trimester before birth. Thereafter, the number of MNs remains relatively stable, with their connections to skeletal muscle dependent on neurotrophins, including brain-derived neurotrophic factor (BDNF) signaling through its high-affinity full-length tropomyosin-related kinase receptor subtype B (TrkB.FL) receptor. As a genetic knockout of BDNF leads to extensive MN loss and postnatal death within 1-2 days after birth, we tested the hypothesis that postnatal inhibition of BDNF/TrkB.FL signaling is important for postnatal phrenic MN (PhMN) survival. In the present study, we used a 1NMPP1-sensitive TrkBF616A mutant mouse to evaluate the effects of inhibition of TrkB kinase activity on phrenic MN (PhMN) numbers and diaphragm muscle (DIAm) fiber cross-sectional area (CSA). Pups were exposed to 1NMPP1 or vehicle (DMSO) from birth to 21 days old (weaning) via the mother's ingestion in the drinking water. Following weaning, the right phrenic nerve was exposed in the neck and the proximal end dipped in a rhodamine solution to retrogradely label PhMNs. After 24 h, the cervical spinal cord and DIAm were excised. Labeled PhMNs were imaged using confocal microscopy, whereas DIAm strips were frozen at ∼1.5× resting length, cryosectioned, and stained with hematoxylin and eosin to assess CSA. We observed an ∼34% reduction in PhMN numbers and increased primary dendrite numbers in 1NMPP1-treated TrkBF616A mice. The distribution of PhMN size (somal surface area) DIAm fiber cross-sectional areas did not differ. We conclude that survival of PhMNs during early postnatal development is sensitive to BDNF/TrkB.FL signaling.NEW & NOTEWORTHY During early postnatal development, BDNF/TrkB signaling promotes PhMN survival. Inhibition of BDNF/TrkB signaling in early postnatal development does not impact PhMN size. Inhibition of BDNF/TrkB signaling in early postnatal development does not impact the number or CSA of DIAm fibers.