bluuebunny/crossref_metadata_2025 huggingface dataset:
Opposing Evolutionary Pressures Drive Clonal Evolution and Health Outcomes in the Aging Blood System
Authors: Skead Kimberly , Ang Houle Armande , Abelson Sagi , Fave Marie-Julie , Lin Boxi , Soave David , Wright Stephen , Dick John E. , Morris Quaid , Awadalla Phillip
Published: February 2021

The age-associated accumulation of somatic mutations and large-scale structural variants (SVs) in the early hematopoietic hierarchy have been linked to premalignant stages for cancer and cardiovascular disease (CVD). However, only a small proportion of individuals harboring these mutations progress to disease, and mechanisms driving the transformation to malignancy remains unclear. Hematopoietic evolution, and cancer evolution more broadly, has largely been studied through a lens of adaptive evolution and the contribution of functionally neutral or mildly damaging mutations to early disease-associated clonal expansions has not been well characterised despite comprising the majority of the mutational burden in healthy or tumoural tissues. Through combining deep learning with population genetics, we interrogate the hematopoietic system to capture signatures of selection acting in healthy and pre-cancerous blood populations. Here, we leverage high-coverage sequencing data from healthy and pre-cancerous individuals from the European Prospective Investigation into Cancer and Nutrition Study (n=477) and dense genotyping from the Canadian Partnership for Tomorrow's Health (n=5,000) to show that blood rejects the paradigm of strictly adaptive or neutral evolution and is subject to pervasive negative selection. We observe clear age associations across hematopoietic populations and the dominant class of selection driving evolutionary dynamics acting at an individual level. We find that both the location and ratio of passenger to driver mutations are critical in determining if positive selection acting on driver mutations is able to overwhelm regulated hematopoiesis and allow clones harbouring disease-predisposing mutations to rise to dominance. Certain genes are enriched for passenger mutations in healthy individuals fitting purifying models of evolution, suggesting that the presence of passenger mutations in a subset of genes might confer a protective role against disease-predisposing clonal expansions. Finally, we find that the density of gene disruption events with known pathogenic associations in somatic SVs impacts the frequency at which the SV segregates in the population with variants displaying higher gene disruption density segregating at lower frequencies. Understanding how blood evolves towards malignancy will allow us to capture cancer in its earliest stages and identify events initiating departures from healthy blood evolution. Further, as the majority of mutations are passengers, studying their contribution to tumorigenesis, will unveil novel therapeutic targets thus enabling us to better understand patterns of clonal evolution in order to diagnose and treat disease in its infancy.

Disclosures

Dick: Bristol-Myers Squibb/Celgene: Research Funding.
Integration of EHR and Cancer Registry Data to Construct a Childhood Cancer Survivorship Cohort to Improve Long-Term Follow-up Care for Leukemia and Lymphoma Survivors
Authors: Noyd David H. , Neely Nigel B. , Howell Claire , Oeffinger Kevin C , Kreissman Susan
Published: February 2021

Background and aims:

Marked improvements in pediatric oncology care for children with acute lymphoblastic leukemia (ALL) and leukemia emphasize the importance of appropriate follow-up care for childhood cancer survivors (CCS) to monitor for late effects. The electronic health record (EHR) sparks unique approaches to construct, maintain, and leverage CCS cohorts to improve care. The primary purpose of this study is to utilize a novel approach to integrate EHR and cancer registry data to construct a CCS cohort for ALL and lymphoma survivors. Data mining in the EHR facilitates stratification of patients into risk cohorts and identification of high risk patients with inadequate subspecialty survivorship care

Methods

Cancer registry data from January 1, 1994 to November 30, 2012 provided a base cohort from which pediatric oncology clinic visits were extracted from the EHR. Explanatory variables included gender, race/ethnicity, risk stratification, distance to the medical center, rural-urban commuting area code, and area deprivation index as a measure of socioeconomic status. Primary outcomes included date of last clinic visit to determine appropriate follow-up, defined as being seen in a subspecialty clinic between five and seven years after initial diagnosis. Patients who died or relapsed during the seven year follow-up period were excluded from the analysis.

Results

Between January 1, 1994 and November 30, 2012, there were 262 pediatric oncology patients evaluated at our institution and reported in the cancer registry who were alive and without evidence of recurrence seven years after initial diagnosis. Of these patients, 22% of patients (n=57) were considered to have inadequate follow-up care. In univariate analysis, younger age (p<0.001), primary diagnosis of ALL compared to lymphoma (p=0.007), low risk strata (p=0.014) and closer proximity to primary treatment center or Children's Oncology Group-affiliate site (p=0.013 and 0.026, respectively) were associated with a higher likelihood of follow-up care. Multivariable logistic regression modeling is currently ongoing.

Conclusions

Integration of EHR and cancer registry data represents a feasible and novel approach to construct a cohort of childhood ALL and lymphoma survivors, risk stratify patients based on treatment exposures, and assess adequate subspecialty survivorship care. Future applications include refinement of treatment exposures through the EHR, adherence to guideline recommendations, and other late effects outcomes.

Disclosures

No relevant conflicts of interest to declare.
Mdm2 Loss of Heterozygosity Cooperates with Mutant p53 in Acute Myeloid Leukemia
Authors: Pourebrahim Rasoul , Montoya Rafael Heinz , Alaniz Zoe , Ostermann Lauren B , Luong Hung Alex , Khoury Joseph D. , Andreeff Michael
Published: February 2021

TP53 mutations are observed in 5 to10% of de novo AML cases with dismal disease prognosis and response to therapy. We found that MDM2 loss of heterozygosity (MDM2 LOH) in AML was always concomitant with TP53 missense mutations (log2 odds ratio>3, p<.001), and not TP53 deletions or truncations whereas in lymphomas, MDM2 LOH and TP53 mutations were mutually exclusive. To understand the functional association of MDM2 LOH and TP53 mutation in AML, we generated a conditional somatic Trp53 mutant AML mouse by combining a Trp53R172H mutation with Mdm2 haploinsufficiency in hematopoietic cells using Vav-cre transgenic mice combined with RosamTmG (mTmG) allele as tracing reporter. The conditional mutant Trp53 allele used in this study harbored a R172H mutation downstream of a wildtype Trp53 cDNA flanked by loxP sites (Trp53wm-R172H) so that the expression of the wildtype Trp53 was preserved before the recombination. Mouse cohorts were generated to harbor one Trp53wm-R172H allele and a Trp53fl or were homozygous for the floxed Trp53 allele (Trp53fl/fl). The expression of Cre in hematopoietic stem cells resulted in heterozygous deletion of Mdm2 and conversion of WT Trp53 to Trp53R172, which corresponds to TP53R175H, one of the most frequent mutations in AML. Mice with somatic Trp53 mutation died due to lymphoma, however, Mdm2 haploinsufficient mice combined with Trp53 mutation succumbed to AML. The disease was immunophenotypically and histologically characterized as AML based on the bone marrow histopathology and expression of Cd11b and Ly6A on the surface of leukemic cells. Immunohistopathology of bone marrow and spleen of MdmLOH p53Mut showed a highly proliferative AML characterized by strong nuclear expression of Ki67 and mutant p53 in the bone marrow of mice. Together, our data identifies Mdm2 LOH to be essential for malignant transformation of p53 mutant myeloid lineage leading to AML development.

Disclosures

Andreeff: Daiichi-Sankyo; Breast Cancer Research Foundation; CPRIT; NIH/NCI; Amgen; AstraZeneca: Research Funding; Daiichi-Sankyo; Jazz Pharmaceuticals; Celgene; Amgen; AstraZeneca; 6 Dimensions Capital: Consultancy; Amgen: Research Funding; Centre for Drug Research & Development; Cancer UK; NCI-CTEP; German Research Council; Leukemia Lymphoma Foundation (LLS); NCI-RDCRN (Rare Disease Clin Network); CLL Founcdation; BioLineRx; SentiBio; Aptose Biosciences, Inc: Membership on an entity's Board of Directors or advisory committees.
COVID-19 and Hematologic Malignancies: Single Institution Analysis
Authors: Anusim Nwabundo , Smalley Melanie , Zimmer Markie Sue , Lang Doyle , Jindal Vishal , Ahaneku Hycienth O , Gupta Ruby , Franklin Candace , Jaiyesimi Ishmael
Published: February 2021

Introduction

COVID-19 in patients with hematologic malignancies are poorly characterized. Theoretically, it is believed that patients with malignancies having a significant chance of succumbing to COVID-19 due to the immunosuppression from the disease and myelosuppression from the cancer therapy compared to their counterparts without malignancies. However, there is scant evidence to support this claim. Here we analyze the outcomes of patients with hematologic malignancies and COVID-19.

Methods

This is a retrospective review of all patients with hematologic malignancies with confirmed COVID-19 admitted to any one of the Beaumont hospitals in Michigan between March 1st,2020 and July 1st ,2020. Data regarding the demographics, cancer diagnosis and management, covid-19 management and outcomes were abstracted.

Results

A total of 32 patients with hematologic malignancies were identified. Of these 32 patients, five patients also had solid malignancies. Median age was 70.5 years (22-91 years) with patients being predominantly female 18 (56 %) and Caucasians 16 (50%). Median BMI was 26.5 kg/m2 (15-79 kg/m2).

The most prevalent hematologic malignancies were Chronic/Small Lymphocytic lymphoma 7(25%) and Multiple Myeloma 6(19%). The median length of stay in the hospital was 9 days (2-61days).

Twelve patients were on active therapy prior to COVID-19 diagnosis. Therapies administered include radiation, surgery, anti-CD30 antibody drug conjugate, immunomodulatory agents, proteasome inhibitors, Bruton tyrosine kinase inhibitors, steroids and chemotherapy. These agents were held at the time of diagnosis of COVID-19

Twenty two patients (69%) patients received a combination of hydroxychloroquine and azithromycin which were standard of care at that time. A total of 31 (97%) patients were on anticoagulation of which 14 (45%) were on therapeutic doses while 17 (55%) were on prophylactic dose.

Four (13%) of the patients were admitted in the ICU. A total of 6 deaths were reported and the median age was 83 years (66-91 years). Four deaths were attributed to COVID-19. Two non-COVID-19 deaths were as a result of aspiration pneumonia and hospital acquired pneumonia. Only one patient with COVID-19 received treatment for malignancy (surgical cytoreduction) but succumbed to COVID-19 four days later. All patients who died received a combination of hydroxychloroquine and azithromycin.

Conclusion

Our study, though limited by sample size, shows that about a third of patients received treatment for their hematologic malignancies around the time of COVID-19 diagnosis and only one death was reported. Patients are currently being advised to participate in clinical trials to better evaluate outcomes while on treatment for hematologic malignancies.

Disclosures

No relevant conflicts of interest to declare.
Therapeutic Cancer Vaccination Targeting Shared Tumor Associated Antigens in Combination with Azacitidine for High Risk Myelodysplastic Syndrome - a Phase I Clinical Trial
Authors: Holmberg-Thydén Staffan , Dufva Inge Høgh , Ortved Gang Anne , Fredslund Breinholt Marie , Schejbel Lone , Andersen Mette Klarskov , Kadivar Mohammad , Svane Inge Marie , Grønbæk Kirsten , Reker Hadrup Sine , El Fassi Daniel
Published: February 2021

Background: Standard care for patients with high risk myelodysplastic syndrome (MDS) is hypomethylating agents, such as azacitidine (AZA). AZA can induce expression of silenced genes, including methylated tumor associated antigens. Such tumor associated antigens may be recognized by T cells, and therefore exploited for immunotherapeutic targeting. To our knowledge, this is the first clinical study that combine hypomethylating agents with a multi-peptide therapeutic cancer vaccine in a hematological malignancy.

Method: In this open label phase 1 trial (ClinicalTrials.gov NCT02750995), we combine AZA with a peptide vaccine targeting antigens encoded from NY-ESO-1, MAGE-A3, PRAME and WT-1, which have previously been demonstrated to be upregulated by AZA treatment. Four long synthetic peptides containing previously described class I and class II epitopes for a variety HLA types was emulsified in Incomplete Freund's Adjuvant for subcutaneous injection. Patients were included following verified treatment response to six courses of AZA monotherapy.

Result: Five patients were included in the study and treated with the vaccine. The combination therapy showed only few adverse events during the study period, whereof none classified as serious. There was one instance of grade 4 toxicity; a case of neutrophil count decrease, requiring administration of prophylactic antibiotics, and two instances of grade 3 toxicity; platelet count decrease and neutrophil count decrease. No vaccine-specific immune response could be detected using intracellular cytokine staining or ELISpot assays, however changes in the phenotypic composition of immune cells and their expression of stimulatory and inhibitory markers were identified in individual patients. All patients progressed to AML with a mean time to progression from inclusion (TTP) of 4.9 months (range 2.8 to 7.6). Survival was 17 months (range 10.9 to 30.6) from MDS diagnosis. Sequencing of bone marrow showed clonal evolution of malignant cells, as well as appearance of novel mutations.

Conclusion: The patients progressed to AML with an average time of only five months after initiating the combination therapy. This may be unrelated to the experimental treatment, but the trial was terminated early as there was no sign of clinical benefit or immunological response.

Figure 1. (a) Trial design. All participants received six courses of AZA prior to inclusion and were evaluated with bone marrow biopsy for treatment response. Vaccination was given together with the next three courses of AZA. (b) Vaccine composition. Synthetic long peptides from NY-ESO-1, PRAME, MAGE-A3 and WT-1 were emulsified in adjuvant Montanide ISA 51.

Figure 1

Disclosures

No relevant conflicts of interest to declare.
Outcomes in Patients with Hematological Malignancies Receiving Anti-CD20 Therapy in the Setting of COVID-19 Infection
Authors: Lopez Carlos A , Chitty David , Chi Jeffrey , Saif Wasif , Goldberg Bradley
Published: February 2021

Objective: The global COVID-19 pandemic has posed a major challenge for patients with hematological malignancies undergoing treatment. For patients with acute leukemia and aggressive lymphomas, delaying curative chemotherapy treatments has historically been associated with adverse outcomes. In the absence of any quality prospective data, expert recommendation has been to treat when possible. Though earlier studies have reported that cancer patients are at an increased risk for mortality due to COVID-19 when compared to the general population, questions remain unanswered as to how specific agents used to treat malignancies affect the natural history of COVID-19. In particular, the effect of anti-CD20 therapy in patients with lymphoid malignancies remains unexplored in patients who contract COVID-19 during their treatment.

Methods: We conducted a retrospective chart review of patients at our New York-based institution to identify any COVID-19 positive patients that required inpatient treatment for their lymphoid malignancy from December 31st, 2019 to July 31st, 2020. Only patients that had received anti-CD20 therapy (rituximab or obinutuzumab) were included in this study. COVID-19 positivity was determined via a positive PCR-based test (either nasopharyngeal swab or bronchoalveolar lavage).

Results: A total of 13 patients were identified, with primary hematological diagnoses including DLBCL (6), ALL (1), MALT lymphoma (1), follicular lymphoma (2), and chronic lymphocytic leukemia (1). The COVID-19 related mortality rate in this cohort was 46% (6), with the remainder recovering from COVID-19. Of the 7 patients who survived, 2 patients had persistently positive COVID-19 PCR-based testing after clinical recovery, 2 failed to develop detectable IgG anti-COVID-19 antibodies, 1 had not had antibody testing, and 2 had detectable antibodies. Of the 2 patients that developed antibodies after recovery, 1 had undetectable anti-COVID-19 IgG levels 55 days later on subsequent testing.

Conclusions: Patients who are hospitalized with lymphoid malignancies and are requiring anti-CD20 therapy seem to be at increased risk of COVID-19-related mortality. Furthermore, anti-CD20 therapy appears to adversely impact the development of IgG antibodies and appears to be associated with delayed clearance of the virus on PCR-based testing.

Disclosures

Saif: Lexicon: Research Funding; Yivia: Research Funding; NGM Bio.: Research Funding; Nouscom: Research Funding; ITA Group: Other: Teaching Engagement, Travel Compesation; UpToDate: Patents & Royalties: Royalties; Merck Sharp & Dohme Corporation: Honoraria; Incyte: Research Funding.
Pegfilgrastim Versus Filgrastim in the Supportive Care of Heavily Pretreated Multiple Myeloma in Treatment with Pomalidomide-Dexamethasone
Authors: Cerchione Claudio , Catalano Lucio , Nappi Davide , Pareto Anna Emanuele , Musuraca Gerardo , Lucchesi Alessandro , Pane Fabrizio , Martinelli Giovanni
Published: February 2021

Background

Pegfilgrastim is a pegylated long-acting recombinant form of G-CSF that extends the half-life and allows for once-per-cycle dosing, requiring less frequent dosing than nonpegylated G-CSF.

Aims

The objective of this study was to compare the efficacy and safety of pegfilgrastim in patients affected by heavily pretreated MM, treated with pomalidomide-dexamethasone, in order to determine whether a single subcutaneous injection of pegfilgrastim is as effective as daily injections of standard filgrastim, in terms of haematological toxicity, febrile neutropenic episodes, antibiotic usage and hospedalization duration.

Methods

57 patients (31 M and 26 F) were enrolled, median age at diagnosis 69 years (r. 52-84), and median age at start of treatment 76 years (r.56-90) treated with several lines of treatments (median 7, r. 2-12), every refractory to all the drugs previously received, received Pomalidomide-Dexamethasone (P 4 mg for 21 days, D 40 mg days 1,8,15,22, pegfilgrastim day +8) every 28 days, until progression.

Results

Since first course, received in domestic setting, with a very good compliance, patients performed blood counts once weekly and received, from day +8 to day +19, prophylactic oral chinolonic antibiotics and anti-fungal drugs. During neutropenia after first cycle, Filgrastim (5 μgr/kg/day for 3 days) was given if neutrophils count was <1500 x 10^9 cells/L. Median number of filgrastim administrations was 4.6 (r. 3-6); nadir neutropenia was registered after a median of 10.4 days (r. 7-14); median of nadir neutrophil count was 1.13 x 10^9 cells/L (r.0.3 - 1.5), with maximum duration of 14 days. From the second course, all patients switched to prophylaxis with pegfilgrastim (6 mg), injected subcutaneously with a single administration on day +3 independently from the neutrophil count at that time. During pegfilgrastim, neutropenia was never longer than 8 days, with a consequent reduction of neutropenia-related infections. Median nadir neutrophil count, evaluated for every patients for at least three courses of therapy (r. 3-6) registered at day +11, was 1.28 (r.0.9-2.2). Only 4 patients needed a supplement of 3 administrations of filgrastim. Pegfilgrastim was well tolerated in all patients: main side effects in our patients were mild fever and bone pain (21.2%).

Conclusions

In patients affected by heavily pretreated MM treated with pomalidomide-dexamethasone, pegfilgrastim seems to reduce the incidence of severe neutropenia and infections and may increase the possibility to maintain the scheduled time of treatment.

Disclosures

Lucchesi: Novartis: Honoraria; Incyte: Honoraria; Pfizer: Honoraria.
Validating a Novel Framework to Classify Inpatient Hematology Consultation Requests
Authors: Patell Rushad , Forbath Natalia , Dodge Laura , Carney Brian J. , Cool Josephine , Persaud Brian , Ricotta Daniel N , Hale Andrew J , Freed Jason A
Published: February 2021

Background:

Consistent classification of consult requests may lead to more productive, efficient, and collegial conversations about patient care, which can facilitate improved work satisfaction and an enhanced learning environment. We propose an organizing framework of 7 specific consultation types: ideal, obligatory, procedural, S.O.S., confirmatory, inappropriate, and curbside. We aimed to obtain validity evidence for this rubric to consistently classify consultation requests in an academic setting.

Methods:

A random sample of 100 de-identified hematology oncology consultation requests made through the online consult portal from a single academic center were selected and independently coded as 1 of the 7 consultation types by 3 hematologists and 3 hospitalists. Perfect (same consult assignment by all coders) and partial (same consult assignment by >4/6 coders) concordance was calculated. Perfect and partial (>2/3 coders) inter-rater concordance based on consult subtypes and provider specialty was also calculated. To assess if length of consult request has an impact on the classification of consult, the length of the request was compared across concordant, partially concordant, and discordant requests.

Results:

Of the 100 consults, perfect concordance was 57%, and partial concordance was 92% (Figure 1). Perfect concordance was 69% amongst hematologists and 78% amongst hospitalists. In cases without perfect concordance (n=43), hematologists agreed with each other 76% of the time, while hospitalists agreed with each other 81% of the time. Of the consults that at least 4 coders classified in the same way, hospitalists were more likely than hematologists to have perfect concordance for ideal consults (89% vs. 66%, respectively; P=0.001) but less likely to have perfect concordance for S.O.S consults (56% vs. 100%, respectively; P=0.003). Hematologists were twice more likely than hospitalists to classify a consult request as S.O.S (26.7% vs. 12.3%, P<0.001), while a greater proportion of hospitalists classified consults as ideal (74% vs. 61%, P=0.007). There was no significant difference in the word count of requests that were perfectly concordant (68+35), partially concordant (65+51), or discordant (39+22) (P=0.18).

Conclusion:

Hematology oncology consult requests can be classified into a novel rubric of 7 specific subtypes. Overall, partial concordance between primary providers and consultants was high, and perfect concordance was moderate. Hematologists were more likely to classify consult requests as S.O.S (without specific questions) than primary providers. Opportunities exist to utilize the rubric to improve communication between health care providers and to improve the medical education of trainees.

Disclosures

No relevant conflicts of interest to declare.
A zebrafish model of granulin deficiency reveals essential roles in myeloid cell differentiation
Authors: Campbell Clyde A. , Fursova Oksana , Cheng Xiaoyi , Snella Elizabeth , McCune Abbigail , Li Liangdao , Solchenberger Barbara , Schmid Bettina , Sahoo Debashis , Morton Mark , Traver David , Espín-Palazón Raquel
Published: February 2021

Granulin is a pleiotropic protein involved in inflammation, wound healing, neurodegenerative disease, and tumorigenesis. These roles in human health have prompted research efforts to use granulin to treat rheumatoid arthritis and frontotemporal dementia and to enhance wound healing. But how granulin contributes to each of these diverse biological functions remains largely unknown. Here, we have uncovered a new role for granulin during myeloid cell differentiation. We have taken advantage of the tissue-specific segregation of the zebrafish granulin paralogues to assess the functional role of granulin in hematopoiesis without perturbing other tissues. By using our zebrafish model of granulin deficiency, we revealed that during normal and emergency myelopoiesis, myeloid progenitors are unable to terminally differentiate into neutrophils and macrophages in the absence of granulin a (grna), failing to express the myeloid-specific genes cebpa, rgs2, lyz, mpx, mpeg1, mfap4, and apoeb. Functionally, macrophages fail to recruit to the wound, resulting in abnormal healing. Our CUT&RUN experiments identify Pu.1, which together with Irf8, positively regulates grna expression. In vivo imaging and RNA sequencing experiments show that grna inhibits the expression of gata1, leading to the repression of the erythroid program. Importantly, we demonstrated functional conservation between the mammalian granulin and the zebrafish ortholog grna. Our findings uncover a previously unrecognized role for granulin during myeloid cell differentiation, which opens a new field of study that can potentially have an impact on different aspects of human health and expand the therapeutic options for treating myeloid disorders such as neutropenia or myeloid leukemia.
PML-controlled responses in severe congenital neutropenia withELANE-misfolding mutations
Authors: Olofsen Patricia A. , Bosch Dennis A. , Roovers Onno , van Strien Paulina M. H. , de Looper Hans W. J. , Hoogenboezem Remco M. , Barnhoorn Sander , Mastroberardino Pier G. , Ghazvini Mehrnaz , van der Velden Vincent H. J. , Bindels Eric M. J. , de Pater Emma M. , Touw Ivo P.
Published: February 2021

Mutations in ELANE cause severe congenital neutropenia (SCN), but how they affect neutrophil production and contribute to leukemia predisposition is unknown. Neutropenia is alleviated by CSF3 (granulocyte colony-stimulating factor) therapy in most cases, but dose requirements vary between patients. Here, we show that CD34+CD45+ hematopoietic progenitor cells (HPCs) derived from induced pluripotent stem cell lines from patients with SCN that have mutations in ELANE (n = 2) or HAX1 (n = 1) display elevated levels of reactive oxygen species (ROS) relative to normal iPSC-derived HPCs. In patients with ELANE mutations causing misfolding of the neutrophil elastase (NE) protein, HPCs contained elevated numbers of promyelocyte leukemia protein nuclear bodies, a hallmark of acute oxidative stress. This was confirmed in primary bone marrow cells from 3 additional patients with ELANE-mutant SCN. Apart from responding to elevated ROS levels, PML controlled the metabolic state of these ELANE-mutant HPCs as well as the expression of ELANE, suggestive of a feed-forward mechanism of disease development. Both PML deletion and correction of the ELANE mutation restored CSF3 responses of these ELANE-mutant HPCs. These findings suggest that PML plays a crucial role in the disease course of ELANE-SCN characterized by NE misfolding, with potential implications for CSF3 therapy.
Influence of height on endothelial maintenance activity: a narrative review
Authors: Shimizu Yuji , Maeda Takahiro
Published: February 2021

Recent studies have revealed an inverse association between height and cardiovascular disease. However, the background mechanism of this association has not yet been clarified. Height has also been reported to be positively associated with cancer. Therefore, well-known cardiovascular risk factors, such as increased oxidative stress and chronic inflammation, are not the best explanations for this inverse association because these risk factors are also related to cancer. However, impaired blood flow is the main pathological problem in cardiovascular disease, while glowing feeding vessels (angiogenesis) are the main characteristic of cancer pathologies. Therefore, endothelial maintenance activity, especially for the productivity of hematopoietic stem cells such as CD34-positive cells, could be associated with the height of an individual because this cell contributes not only to the progression of atherosclerosis but also to the development of angiogenesis. In addition, recent studies have also revealed a close connection between bone marrow activity and endothelial maintenance; bone marrow-derived hematopoietic stem cells contribute towards endothelial maintenance. Since the absolute volume of bone marrow is positively associated with height, height could influence endothelial maintenance activity. Based on these hypotheses, we performed several studies. The aim of this review is not only to discuss the association between height and bone marrow activity, but also to describe the potential mechanism underlying endothelial maintenance. In addition, this review also aims to explain some of the reasons that implicate hypertension as a major risk factor for stroke among the Japanese population. The review also aims to clarify the anthropological reasons behind the high risk of atherosclerosis progression in Japanese individuals with acquired genetic characteristics.
Genomics Analyses Reveal Unique Classification, Population Structure and Novel Allele of Neo-Tetraploid Rice
Authors: Yu Hang , Li Qihang , Li Yudi , Yang Huijing , Lu Zijun , Wu Jinwen , Zhang Zemin , Shahid Muhammad Qasim , Liu Xiangdong
Published: February 2021

Background

Neo-tetraploid rice (NTR) is a useful new germplasm that developed from the descendants of the autotetraploid rice (ATR) hybrids. NTR showed improved fertility and yield potential, and produced high yield heterosis when crossed with indica ATR for commercial utilization. However, their classification, population structure and genomic feature remain elusive.

Results

Here, high-depth genome resequencing data of 15 NTRs and 18 ATRs, together with 38 publicly available data of diploid rice accessions, were analyzed to conduct classification, population structure and haplotype analyses. Five subpopulations were detected and NTRs were clustered into one independent group that was adjacent to japonica subspecies, which maybe the reason for high heterosis when NTRs crossed with indica ATRs. Haplotype patterns of 717 key genes that associated with yield and other agronomic traits were revealed in these NTRs. Moreover, a novel specific SNP variation was detected in the first exon of HSP101, a known heat-inducible gene, which was conserved in all NTRs but absent in ATRs, 3KRG and RiceVarMap2 databases. The novel allele was named as HSP101–1, which was confirmed to be a heat response factor by qRT-PCR, and knockout of HSP101–1 significantly decreased the thermotolerance capacity of NTR. Interestingly, HSP101–1 was also specifically expressed in the anthers of NTR at pre-meiotic and meiosis stages under optimal environment without heat stress, and its loss-of-function mutant showed significant decrease in fertility of NTR.

Conclusion

The construction of first genomic variation repository and the revelation of population structure provide invaluable information for optimizing the designs of tetraploid rice breeding. The detection of specific genomic variations offered useful genomic markers and new directions to resolve high fertility mechanism of NTR.
Quantum dots as targeted doxorubicin drug delivery nanosystems in human lung cancer cells
Authors: Ruzycka-Ayoush Monika , Kowalik Patrycja , Kowalczyk Agata , Bujak Piotr , Nowicka Anna M. , Wojewodzka Maria , Kruszewski Marcin , Grudzinski Ireneusz P.
Published: February 2021

Background

Lung cancer is one of the most frequently diagnosed cancers all over the world and is also one of the leading causes of cancer-related mortality. The main treatment option for small cell lung cancer, conventional chemotherapy, is characterized by a lack of specificity, resulting in severe adverse effects. Therefore, this study aimed at developing a new targeted drug delivery (TDD) system based on Ag–In–Zn–S quantum dots (QDs). For this purpose, the QD nanocrystals were modified with 11-mercaptoundecanoic acid (MUA), L-cysteine, and lipoic acid decorated with folic acid (FA) and used as a novel TDD system for targeting doxorubicin (DOX) to folate receptors (FARs) on adenocarcinomic human alveolar basal epithelial cells (A549). NIH/3T3 cells were used as FAR-negative controls. Comprehensive physicochemical, cytotoxicity, and genotoxicity studies were performed to characterize the developed novel TDDs.

Results

Fourier transformation infrared spectroscopy, dynamic light scattering, and fluorescence quenching confirmed the successful attachment of FA to the QD nanocrystals and of DOX to the QD–FA nanocarriers. UV–Vis analysis helped in determining the amount of FA and DOX covalently anchored to the surface of the QD nanocrystals. Biological screening revealed that the QD–FA–DOX nanoconjugates had higher cytotoxicity in comparison to the other forms of synthesized QD samples, suggesting the cytotoxic effect of DOX liberated from the QD constructs. Contrary to the QD–MUA–FA–DOX nanoconjugates which occurred to be the most cytotoxic against A549 cells among others, no such effect was observed for NIH/3T3 cells, confirming FARs as molecular targets. In vitro scratch assay also revealed significant inhibition of A549 cell migration after treatment with QD–MUA–FA–DOX. The performed studies evidenced that at IC50 all the nanoconjugates induced significantly more DNA breaks than that observed in nontreated cells. Overall, the QD–MUA–FA–DOX nanoconjugates showed the greatest cytotoxicity and genotoxicity, while significantly inhibiting the migratory potential of A549 cells.

Conclusion

QD–MUA–FA–DOX nanoconjugates can thus be considered as a potential drug delivery system for the effective treatment of adenocarcinomic human alveolar basal epithelial cells.
Targeting BMI-1 with PLGA–PEG nanoparticle-containing PTC209 modulates the behavior of human glioblastoma stem cells and cancer cells
Authors: Poonaki Elham , Ariakia Fatemeh , Jalili-Nik Mohammad , Shafiee Ardestani Mehdi , Tondro Gholamhossein , Samini Fariborz , Ghasemi Sepideh , Sahab-Negah Sajad , Gorji Ali
Published: February 2021

Despite advances in glioblastoma (GBM) treatments, current approaches have failed to improve the overall survival of patients. The oncogene BMI-1, a core member of the polycomb group proteins, is a potential novel therapeutic target for GBM. To enhance the efficacy and reduce the toxicity, PTC209, a BMI-1 inhibitor, was loaded into a PLGA–PEG nanoparticle conjugated with CD133 antibody (Nano-PTC209) and its effect on the behavior of human GBM stem-like cells (GSCs) and the human glioblastoma cell line (U87MG) was assessed. Nano-PTC209 has a diameter of ~ 75 nm with efficient drug loading and controlled release. The IC50 values of Nano-PTC209 for GSCs and U87MG cells were considerably lower than PTC209. Nano-PTC209 significantly decreased the viability of both GSCs and U87MG cells in a dose-dependent manner and caused a significant enhancement of apoptosis and p53 levels as well as inhibition of AKT and JNK signaling pathways. Furthermore, Nano-PTC209 significantly inhibited the migration ability, decreased the activity of metalloproteinase-2 and -9, and increased the generation of reactive oxygen species in both GSCs and U87MG cells. Our data indicate that PLGA–PEG nanoparticle conjugated with CD133 antibody could be an ideal nanocarrier to deliver PTC209 and effectively target BMI-1 for potential approaches in the treatment of GBM.
Non-invasive transdermal delivery of chemotherapeutic molecules in vivo using superparamagnetic iron oxide nanoparticles
Authors: Raviraj Vanisri , Pham Binh T. T. , Kim Byung J. , Pham Nguyen T. H. , Kok Lai F. , Painter Nicole , Delic Naomi C. , Jones Stephen K. , Hawkett Brian S. , Lyons J. Guy
Published: February 2021

Background

The skin is both a target and a potential conduit for the delivery of drugs, but its cornified cell layer resists penetration by most molecules. This study investigated the potential of superparamagnetic iron oxide nanoparticles to facilitate the transdermal delivery of anticancer agents.

Results

Chemotherapeutic cancer drugs were applied with or without nanoparticles to the skin of hairless mice, and their ability to penetrate the skin was assessed using fluorescence microscopy and tumor growth. Nanoparticles enhanced the penetration of the skin by doxorubicin and 5-fluorouracil as determined by fluorescence microscopy and growth retardation of experimental melanoma in immunocompetent, syngeneic mice. This drug enhancement did not require conjugation or encapsulation of the drugs by the nanoparticles—simple co-administration sufficed. Nanoparticles applied topically to melanomas increased the cytotoxicity and immune cell infiltration induced by co-administered 5-fluorouracil, and also reduced vascularization of the tumors independently of 5-fluorouracil.

Conclusion

Correctly formulated superparamagnetic iron oxide nanoparticles can facilitate the chemotherapeutic effectiveness of cytotoxic drugs on skin tumors by both increasing their transdermal penetration and ameliorating host–tumor interactions. This enhancement of skin penetration occurs without the need for conjugation or encapsulation of the co-administered drugs, and so will likely be applicable to other drugs, also.
Investigation of ancestral alleles in the Bovinae subfamily
Authors: Naji Maulana M. , Utsunomiya Yuri T. , Sölkner Johann , Rosen Benjamin D. , Mészáros Gábor
Published: February 2021

Background

In evolutionary theory, divergence and speciation can arise from long periods of reproductive isolation, genetic mutation, selection and environmental adaptation. After divergence, alleles can either persist in their initial state (ancestral allele - AA), co-exist or be replaced by a mutated state (derived alleles -DA). In this study, we aligned whole genome sequences of individuals from the Bovinae subfamily to the cattle reference genome (ARS.UCD-1.2) for defining ancestral alleles necessary for selection signatures study.

Results

Accommodating independent divergent of each lineage from the initial ancestral state, AA were defined based on fixed alleles on at least two groups of yak, bison and gayal-gaur-banteng resulting in ~ 32.4 million variants. Using non-overlapping scanning windows of 10 Kb, we counted the AA observed within taurine and zebu cattle. We focused on the extreme points, regions with top 0. 1% (high count) and regions without any occurrence of AA (null count). High count regions preserved gene functions from ancestral states that are still beneficial in the current condition, while null counts regions were linked to mutated ones. For both cattle, high count regions were associated with basal lipid metabolism, essential for survival of various environmental pressures. Mutated regions were associated to productive traits in taurine, i.e. higher metabolism, cell development and behaviors and in immune response domain for zebu.

Conclusions

Our findings suggest that retaining and losing AA in some regions are varied and made it species-specific with possibility of overlapping as it depends on the selective pressure they had to experience.
Geranylgeranylacetone attenuates cerebral ischemia–reperfusion injury in rats through the augmentation of HSP 27 phosphorylation: a preliminary study
Authors: Matsuo Kazuya , Hosoda Kohkichi , Tanaka Jun , Yamamoto Yusuke , Imahori Taichiro , Nakai Tomoaki , Irino Yasuhiro , Shinohara Masakazu , Sasayama Takashi , Kohmura Eiji
Published: February 2021

Background

We previously reported that heat shock protein 27 (HSP27) phosphorylation plays an important role in the activation of glucose-6-phosphate dehydrogenase (G6PD), resulting in the upregulation of the pentose phosphate pathway and antioxidant effects against cerebral ischemia–reperfusion injury. The present study investigated the effect of geranylgeranylacetone, an inducer of HSP27, on ischemia–reperfusion injury in male rats as a preliminary study to see if further research of the effects of geranylgeranylacetone on the ischemic stroke was warranted.

Methods

In all experiments, male Wistar rats were used. First, we conducted pathway activity profiling based on a gas chromatography–mass spectrometry to identify ischemia–reperfusion-related metabolic pathways. Next, we investigated the effects of geranylgeranylacetone on the pentose phosphate pathway and ischemia–reperfusion injury by real-time polymerase chain reaction (RT-PCR), immunoblotting, and G6PD activity, protein carbonylation and infarct volume analysis. Geranylgeranylacetone or vehicle was injected intracerebroventricularly 3 h prior to middle cerebral artery occlusion or sham operation.

Results

Pathway activity profiling demonstrated that changes in the metabolic state depended on reperfusion time and that the pentose phosphate pathway and taurine-hypotaurine metabolism pathway were the most strongly related to reperfusion among 137 metabolic pathways. RT-PCR demonstrated that geranylgeranylacetone did not significantly affect the increase in HSP27 transcript levels after ischemia–reperfusion. Immunoblotting showed that geranylgeranylacetone did not significantly affect the elevation of HSP27 protein levels. However, geranylgeranylacetone significantly increase the elevation of phosphorylation of HSP27 after ischemia–reperfusion. In addition, geranylgeranylacetone significantly affected the increase in G6PD activity, and reduced the increase in protein carbonylation after ischemia–reperfusion. Accordingly, geranylgeranylacetone significantly reduced the infarct size (median 31.3% vs 19.9%, p = 0.0013).

Conclusions

As a preliminary study, these findings suggest that geranylgeranylacetone may be a promising agent for the treatment of ischemic stroke and would be worthy of further study. Further studies are required to clearly delineate the mechanism of geranylgeranylacetone-induced HSP27 phosphorylation in antioxidant effects, which may guide the development of new approaches for minimizing the impact of cerebral ischemia–reperfusion injury.
Cloning and overexpression of PeWRKY31 from Populus × euramericana enhances salt and biological tolerance in transgenic Nicotiana
Authors: Yu Xiaoyue , Pan Yu , Dong Yan , Lu Bin , Zhang Chao , Yang Minsheng , Zuo Lihui
Published: February 2021

Background

As important forest tree species, biological stress and soil salinization are important factors that restrict the growth of Populus × euramericana. WRKYs are important transcription factors in plants that can regulate plant responses to biotic and abiotic stresses. In this study, PeWRKY31 was isolated from Populus × euramericana, and its bioinformation, salt resistance and insect resistance were analyzed. This study aims to provide guidance for producing salt-resistant and insect-resistant poplars.

Results

PeWRKY31 has a predicted open reading frame (ORF) of 1842 bp that encodes 613 amino acids. The predicted protein is the unstable, acidic, and hydrophilic protein with a molecular weight of 66.34 kDa, and it has numerous potential phosphorylation sites, chiefly on serines and threonines. PeWRKY31 is a zinc-finger C2H2 type-II WRKY TF that is closely related to WRKY TFs of Populus tomentosa, and localizes to the nucleus. A PeWRKY31 overexpression vector was constructed and transformed into Nicotiana tabacum L. Overexpression of PeWRKY31 improved the salt tolerance and insect resistance of the transgenic tobacco. Transcriptome sequencing and KEGG enrichment analysis showed the elevated expression of genes related to glutathione metabolism, plant hormone signal transduction, and MAPK signaling pathways, the functions of which were important in plant salt tolerance and insect resistance in the overexpressing tobacco line.

Conclusions

PeWRKY31 was isolated from Populus × euramericana. Overexpression of PeWRKY31 improved the resistance of transgenic plant to salt stress and pest stress. The study provides references for the generation of stress-resistant lines with potentially great economic benefit.
New insights into the role of MADS-box transcription factor gene CmANR1 on root and shoot development in chrysanthemum (Chrysanthemum morifolium)
Authors: Sun Cui-Hui , Wang Jia-Hui , Gu Kai-Di , Zhang Peng , Zhang Xin-Yi , Zheng Cheng-Shu , Hu Da-Gang , Ma Fangfang
Published: February 2021

Background

MADS-box transcription factors (TFs) are the key regulators of multiple developmental processes in plants; among them, a chrysanthemum MADS-box TF CmANR1 has been isolated and described as functioning in root development in response to high nitrate concentration signals. However, how CmANR1 affects root and shoot development remains unclear.

Results

We report that CmANR1 plays a positive role in root system development in chrysanthemum throughout the developmental stages of in vitro tissue cultures. Metabolomics combined with transcriptomics assays show that CmANR1 promotes robust root system development by facilitating nitrate assimilation, and influencing the metabolic pathways of amino acid, glycolysis, and the tricarboxylic acid cycle (TCA) cycle. Also, we found that the expression levels of TFs associated with the nitrate signaling pathways, such as AGL8, AGL21, and LBD29, are significantly up-regulated in CmANR1-transgenic plants relative to the wild-type (WT) control; by contrast, the expression levels of RHD3-LIKE, LBD37, and GATA23 were significantly down-regulated. These results suggest that these nitrate signaling associated TFs are involved in CmANR1-modulated control of root development. In addition, CmANR1 also acts as a positive regulator to control shoot growth and development.

Conclusions

These findings provide potential mechanisms of MADS-box TF CmANR1 modulation of root and shoot development, which occurs by regulating a series of nitrate signaling associated TFs, and influencing the metabolic pathways of amino acid and glycolysis, as well as TCA cycle and nitrate assimilation.
Drought-induced ABA, H2O2 and JA positively regulate CmCAD genes and lignin synthesis in melon stems
Authors: Liu Wei , Jiang Yun , Jin Yazhong , Wang Chenghui , Yang Juan , Qi Hongyan
Published: February 2021

Background

Cinnamyl alcohol dehydrogenase (CAD) is an important enzyme functions at the last step in lignin monomer synthesis pathway. Our previous work found that drought induced the expressions of CmCAD genes and promoted lignin biosynthesis in melon stems.

Results

Here we studied the effects of abscisic acid (ABA), hydrogen peroxide (H2O2) and jasmonic acid (JA) to CmCADs under drought stress. Results discovered that drought-induced ABA, H2O2 and MeJA were prevented efficiently from increasing in melon stems pretreated with fluridone (Flu, ABA inhibitor), imidazole (Imi, H2O2 scavenger) and ibuprofen (Ibu, JA inhibitor). ABA and H2O2 are involved in the positive regulations to CmCAD1, 2, 3, and 5, and JA is involved in the positive regulations to CmCAD2, 3, and 5. According to the expression profiles of lignin biosynthesis genes, ABA, H2O2 and MeJA all showed positive regulations to CmPAL2-like, CmPOD1-like, CmPOD2-like and CmLAC4-like. In addition, positive regulations were also observed with ABA to CmPAL1-like, CmC4H and CmCOMT, with H2O2 to CmPAL1-like, CmC4H, CmCCR and CmLAC17-like, and with JA to CmCCR, CmCOMT, CmLAC11-like and CmLAC17-like. As expected, the signal molecules positively regulated CAD activity and lignin biosynthesis under drought stress. Promoter::GUS assays not only further confirmed the regulations of the signal molecules to CmCAD1~3, but also revealed the important role of CmCAD3 in lignin synthesis due to the strongest staining of CmCAD3 promoter::GUS.

Conclusions

CmCADs but CmCAD4 are positively regulated by ABA, H2O2 and JA under drought stress and participate in lignin synthesis.