Interplay between transcription regulators RUNX1 and FUBP1 activates an enhancer of the oncogene c-KIT and amplifies cell proliferation.

Runt-related transcription factor 1 (RUNX1) is a well-known master regulator of hematopoietic lineages but its mechanisms of action are still not fully understood. Here, we found that RUNX1 localizes on active chromatin together with Far Upstream Binding Protein 1 (FUBP1) in human B-cell precursor lymphoblasts, and that both factors interact in the same transcriptional regulatory complex. RUNX1 and FUBP1 chromatin localization identified c-KIT as a common target gene. We characterized two regulatory regions, at +700 bp and +30 kb within the first intron of c-KIT, bound by both RUNX1 and FUBP1, and that present active histone marks. Based on these regions, we proposed a novel FUBP1 FUSE-like DNA-binding sequence on the +30 kb enhancer. We demonstrated that FUBP1 and RUNX1 cooperate for the regulation of the expression of the oncogene c-KIT. Notably, upregulation of c-KIT expression by FUBP1 and RUNX1 promotes cell proliferation and renders cells more resistant to the c-KIT inhibitor imatinib mesylate, a common therapeutic drug. These results reveal a new mechanism of action of RUNX1 that implicates FUBP1, as a facilitator, to trigger transcriptional regulation of c-KIT and to regulate cell proliferation. Deregulation of this regulatory mechanism may explain some oncogenic function of RUNX1 and FUBP1

Towards the Rational Design of Photoinduced Electron Transfer (PET)-based Fluorescent Probes

Fluorescent probes act as valuable chemical tools to detect a variety of substances with exceptional sensitivity and selectivity. One desired property of an effective fluorescent probe is that it acts via a “turn-on” mechanism, in which fluorescence intensity increases upon interaction with the desired analyte. Photoinduced electron transfer (PET) is one of the many mechanisms that is used to modulate photophysical properties of fluorescent probes. In this mechanism, the fluorogenic probes are equipped with a motif that can suppress fluorescence. These probes can undergo a transformation upon interaction with a target molecule that restricts that moiety from quenching the fluorescence, resulting in a fluorescence enhancement. Various probes containing pendant aryl quenching moieties that undergo photoinduced electron transfer have been reported to have a direct correlation between the experimentally determined quantum yield of the probe and the highest occupied molecular orbital (EHOMO) of the corresponding PET quenching moiety. However, the systematic investigation of the relationship between PET-quenching and moieties other than aryl rings has been underdeveloped. By changing the electronics of heteroatom quenching moieties, systematic studies of factors which affect PET for common fluorophores, including coumarin based systems, can be reported. Through the synthesis of 7-MeO-coumarin probes with various quencher functionality, there exists a correlation between the quantum yield of these probes and the molecular orbital energy of the quenching moiety. This study aims to provide the scientific community with tools to design heteroatom PET-based fluorescent sensors a priori

Functional and clinical consequences of clonal hematopoiesis of indeterminate potential on cardiovascular diseases and cancer

Studien aus den Jahren 2014 und 2015, die Sequenzierungsmethoden der neuesten Generation („next generation sequencing“, NGS) verwendet haben, brachten eine neue Entität hervor, klonale Hämatopoese von unbestimmtem Potenzial („clonal hematopoiesis of indeterminate potential“, CHIP). Sie wird definiert durch das Auftreten von somatischen Mutationen in Genen, die typischerweise in hämatologischen Krankheiten mutiert sind, bei CHIP aber in hämatologischen gesunden Menschen vorkommen. Diese Menschen zeigen ein nur leicht erhöhtes Risiko für spätere Leukämien, aber überraschenderweise eine signifikant verkürzte Lebenserwartung, die hauptsächlich durch ein erhöhtes Risiko für kardiovaskuläre Krankheiten erklärt wird. CHIP ist altersabhängig, sodass 10-20% der Altersgruppe über 70 Jahren CHIP aufweisen. Für CHIP wurde der Schwellenwert mutierter Allele („variant allele frequency“, VAF) auf 2 % definiert. Wegen der Korrelation mit kardiovaskulären Erkrankungen, wurde im Rahmen dieser Dissertation das Knochenmark einer Patientenkohorte (CHF-Kohorte) sequenziert, die an chronischer Herzinsuffizienz litten. Dazu wurde eine sehr genaue Amplikon-basierte NGS-Methode etabliert. Eine Analyse der Blutzellen im Knochenmark der CHF-Kohorte mit TET2-Mutationen hat eine signifikant erhöhte Anzahl von Leukozyten, sowie Stamm- und Vorläuferzellen gezeigt. Die vorliegende Studie konnte außerdem zeigen, dass sich ein signifikant verschlechterten Krankheitsverlauf und eine erhöhte Sterblichkeit ergab, wenn die Patienten eine Mutation in einem der am häufigsten mutierten Genen trugen, DNMT3A oder TET2. Mit der etablierten Sequenziermethode waren auch Mutationen unter einer VAF von 2 % detektierbar. Damit konnte eine Dosis-Abhängigkeit zwischen der Klongröße und klinischem Ausgang der Krankheit bei CHF-Patienten mit CHIP gezeigt werden. Das Risiko für eine kardiovaskuläre Erkrankung und Krebs steigt mit zunehmendem Alter und die Diagnose einer der Krankheiten bedeutet auch ein erhöhtes Risiko, die andere zu entwickeln. Zwischen Krebstherapien und der Entwicklung von kardiovaskulären Veränderungen, sowie sekundären, therapieinduzierten Leukämien, besteht ein oft beobachteter Zusammenhang. Therapieinduzierte Leukämien sind eine relativ häufige Folge von Chemotherapie und folgender autologer Stammzelltransplantation (ASZT). Darum war ein zweiter Schwerpunkt dieser Arbeit eine Patientenkohorte, die mit einer ASZT behandelt wurde. Die meisten Mutationen waren in den transplantierten Zellen zu finden, die dann zahlenmäßig nach der Therapie anstiegen, was für das Vorkommen der Mutationen in den 2 Stamm- und Vorläuferzellen spricht. Eine weitere Schlussfolgerung ist, dass Mutationen einen selektiven Vorteil bei der Wiederherstellung der Blutzellen nach der Therapie haben könnten.Clonal hematopoiesis of indeterminate potential (CHIP) is caused by recurrent somatic mutations leading to clonal blood cell expansion. However, direct evidence of the fitness of CHIP-mutated human hematopoietic stem cells (HSCs) in blood reconstitution is lacking. Because myeloablative treatment and transplantation enforce stress on HSCs, we followed 81 patients with solid tumors or lymphoid diseases undergoing autologous stem cell transplantation (ASCT) for the development of CHIP. We found a high incidence of CHIP (22%) after ASCT with a high mean variant allele frequency (VAF) of 10.7%. Most mutations were already present in the graft, albeit at lower VAFs, demonstrating a selective reconstitution advantage of mutated HSCs after ASCT. Thus, CHIP-mutated stem and progenitor cells largely gain on clone size upon ASCT-related blood reconstitution, leading to an increased future risk of CHIP-associated complications. CHIP increase with age and is also associated with atherosclerosis and inflammation. Age and inflammation are the major risk factors for heart failure, yet the association of CHIP with chronic ischemic heart failure (CHF) in humans is unknown. Therefore, we analyzed bone marrow-derived mononuclear cells from 200 patients with CHF by NGS to detect the presence of CHIP and associated such with long-term prognosis in patients with CHF. Forty-seven mutations with a VAF of at least 2% were found in 18.5% of 200 patients with CHF. The mutations most commonly occurred in the genes DNMT3A and TET2. During a median follow-up of 4.4 years, a significantly worse clinical outcome for patients with either DNMT3A or TET2 mutations compared with non-CHIP carriers was notable. Importantly, there was a significant dose-response association between VAF and clinical outcome. Our data suggest that somatic mutations in hematopoietic cells, may be significantly associated with the progression and poor prognosis of CHF

Toward the Rational Design of Photo-induced Electron Transfer (PET)-Based Fluorescent Probes

Small molecule fluorescent probes are commonly used in imaging applications and sensors including detecting environmental analytes or elucidating biological phenomena. Many of these compounds function via a change in brightness occurring through photoinduced electron transfer (PeT). Although this concept is well understood, PeT-based probes are almost entirely developed via empirical methods. The current project focuses on synthesizing a series of naphthalene and coumarin-based fluorescent probes bearing carefully altered quenching moieties. The photophysical properties of these molecules will then be determined. Ultimately, this systematic investigation of structure-optical property relationship aims to provide the scientific community with a more rational means for creating these useful chemical tools

Preparing Teacher Candidates for Collaborating with Families

Brianna Wright, Childhood Education and Macy Dorsheimer, Early Childhood and Childhood Education Faculty Mentors: Professor Corinne Kindzierski, Elementary Education and Professor Julie Henry, Elementary Education During the spring semester, we observed and researched the teacher education program to gain insight on how teacher candidates can build their confidence working with families. Going into the field, many future educators are apprehensive about parent-teacher conferences or other interactions with diverse families. Due to this lack of knowledge and expertise in the field, many new teachers feel as if they need to continuously practice these skills to better communicate with parents. Our research project goals are to find the best method to address any concerns teacher candidates may have regarding caregiver/parent communication and to identify what advice senior teacher candidates can provide for future teachers. We generated a survey for students in the Education program to identify concerns they may have in working with diverse families and inquire about resources available to them. Using the results, our poster promotes suggestions and other ideas to help future teachers feel more comfortable with working with families. This information will positively impact pre-service teachers and their ability to gain the confidence to acknowledge any issues when collaborating with families.https://digitalcommons.buffalostate.edu/srcc-sp20-edu/1033/thumbnail.jp

Information processing biases: The effects of negative emotional symptoms on sampling pleasant and unpleasant information.

Although theories of emotion associate negative emotional symptoms with cognitive biases in information processing, they rarely specify the details. Here, we characterize cognitive biases in information processing of pleasant and unpleasant information, and how these biases covary with anxious and depressive symptoms, while controlling for general stress and cognitive ability. Forty undergraduates provided emotional symptom scores (Depression Anxiety Stress Scale–21) and performed a statistical learning task that required predicting the next sound in a long sequence of either pleasant or unpleasant naturalistic sounds (blocks). We used an information weights framework to determine if the degree of behavioral change associated with observing either confirmatory (“B” follows “A”) or disconfirmatory (“B” does not follow “A”) transitions differs for pleasant and unpleasant sounds. Bayesian mixed-effects models revealed that negative emotional symptom scores predicted performance as well as processing biases of pleasant and unpleasant information. Further, information weights differed between pleasant and unpleasant information, and importantly, this difference varied based on symptom scores. For example, higher depressive symptom scores predicted a bias of underutilizing disconfirmatory information in unpleasant content. These findings have implications for models of emotional disorders by offering a mechanistic explanation and formalization of the associated cognitive biases

Confocal laser scanning microscopy image analyses of pycnidia of wild and mutant <i>Z</i>. <i>tritici</i>.

<p>WGA-FITC / Propidium iodide double-stained pycnidia on infected wheat leaves 28 dpi of wild-type (A), mutant <i>Z</i>. <i>tritici</i> isolates; IPO323Δ<i>Zt80707</i> (B) and IPO323Δ<i>Zt103264</i> (C). Fungal cell walls (green) and fungal and plant nuclei (red) are visible in the infected plant tissue (scale bar: 50 μm). D) Average size of pycnidia measured 14 and 28 dpi. For each strain and time point, 50 pycnidia were measured. ** <i>p</i><0.01; <i>p</i>-values were calculated using a Mann-Whitney U test. Whiskers indicate standard deviations.</p