Fanconi Anemia Research Articles

Loss of Fanconi anemia proteins causes a reliance on lysosomal exocytosis.

Mutations in the FA pathway lead to a rare genetic disease that increases risk of bone marrow failure, acute myeloid leukemia, and solid tumors. FA patients have a 500 to 800-fold increase in head and neck squamous cell carcinoma compared to the general population and the treatment for these malignancies are ineffective and limited due to the deficiency in DNA damage repair. Using unbiased CRISPR-interference screening, we found the loss of FA function renders cells dependent on key exocytosis genes such as SNAP23. Further investigation revealed that loss of FA pathway function induced deficiencies in lysosomal health, dysregulation of autophagy and increased lysosomal exocytosis. The compromised cellular state caused by the loss of FA genes is accompanied with decreased lysosome abundance and increased lysosomal membrane permeabilization in cells. We found these signatures in vitro across multiple cell types and cell lines and in clinically relevant FA patient cancers. Our findings are the first to connect the FA pathway to lysosomal exocytosis and thus expands our understanding of FA as a disease and of induced dependencies in FA mutant cancers.

Head and Neck Cancer in Fanconi Anemia: Report of 11 Cases and a Systematic Review

Background/Objectives: Head and neck cancer is one of the most common malignancies in patients with Fanconi anemia (FA), with a greater than 500-fold incidence of head and neck squamous cell carcinoma. Treatment of HNSCC in these patients is particularly challenging because of poor tolerance to radiation therapy and to some chemotherapeutic agents such as cisplatin. For these reasons, new research is needed to determine the best treatment course for these patients. With this goal in mind, we assessed the characteristics of head and neck squamous cell carcinoma (SCC) in Fanconi anemia (FA). Subjects and Methods: Data for 11 patients (mean age 31 years) with head and neck SCC and FA attended to between 2005 and 2021 were analyzed. Results: The primary tumor site was the oral cavity in eight patients, and five had advanced stages. All patients underwent primary tumor resection. Four patients received adjuvant radiotherapy (mean 57.2 Gy), but three developed toxicity. The mean follow-up was 48.4 months. Six patients experienced 19 episodes of primary tumor recurrence and five developed secondary head and neck tumors. The 3-year disease-free survival was 47%, and the 5-year cause-specific survival was 48%. These findings are similar to the data for 72 patients collected from the literature. Conclusions: The prognosis of head and neck cancer in patients with Fanconi anemia is poor, with an overall survival lower than 50% at 5 years.

Inhibition of DEK restores hematopoietic stem cell function in Fanconi anemia.

Hematopoietic stem cells (HSCs) are susceptible to replication stress, which is a major contributor to HSC defects in Fanconi anemia (FA). Here, we report that HSCs relax the global chromatin by downregulating the expression of a chromatin architectural protein, DEK, in response to replication stress. DEK is abnormally accumulated in bone marrow (BM) CD34+ cells from patients with FA and in Fancd2-deficient HSCs. DEK haploinsufficiency promotes chromatin relaxation, replication stress relief, and function recovery of Fancd2-deficient HSCs. Furthermore, inhibition of DEK restores the proliferation of FA CD34+ cells in vitro and enhances their engraftment in vivo. Mechanistically, the activating transcription factor 2 (ATF2), specifically phosphorylated ATF2 at Thr69/71, was identified as a promoter of DEK transcription. Fancd2 deficiency results in p38 hyperphosphorylation, which in turn phosphorylates ATF2 at Thr69/71, leading to DEK accumulation in HSCs. In conclusion, our findings establish a functional link between chromatin relaxation and replication stress tolerance in HSCs and highlight DEK as a target for FA.

Phase 1 Study of Quercetin, a Natural Antioxidant in Children and Young Adults with Fanconi Anemia (FA).

Fanconi anemia (FA) is a rare inherited disorder characterized by progressive bone marrow failure (BMF) and a predisposition to malignancy. Systemic reactive-oxygen species (ROS) and increased sensitivity of FA hematopoietic progenitors to ROS play a key role in the pathogenesis of BMF. Treatment with antioxidants improve hematopoietic function in Fancc-/- mice. We report the safety, tolerability, and pharmacokinetics (PK) of quercetin, a naturally occurring antioxidant in the first dose-finding Phase 1 study in patients with FA. Twelve patients (median age 7 years, range: 3-21) received oral quercetin twice daily for 4 months. Quercetin was well tolerated at all dose levels. Allometrically bodyweight-adjusted dose with a maximum adult daily dose of 4000mg/day was established as the recommended dose of quercetin. Patients in an expansion cohort (n=18) were treated using this recommended dose for 6 months. A subset of patients showed reduced ROS levels in the peripheral blood and bone marrow stem cell compartment. Patients in the analysis cohort treated with the recommended dose of quercetin achieved an a priori defined optimal response of 25% reduction in the peripheral blood (PB) ROS level compared to baseline. Platelet counts remained stable to slightly improved over the study period (p=0.06). Absolute neutrophil counts (p=0.01) and hemoglobin levels gradually declined (p=0.001). In those with evidence of BMF at baseline, 8 out of 15 patients (53%) had a hematological response at some point following quercetin treatment. Fluctuations in counts are common in patients with FA limiting accurate assessment of the impact of quercetin use in FA. NCT# 01720147.

Unveiling a pathogenic FANCA gene variant in a Mexican family with Fanconi anemia through next‑generation sequencing.

Fanconi anemia (FA) is the most common hereditary bone marrow failure syndrome, with an incidence of 1 in 5,000,000. This disease is caused by an alteration in one of the 23 genes associated with the FA/BRCA DNA repair pathway, which is responsible for repairing interstrand bridges generated during homologous recombination. FA has been associated with a predisposition to other types of neoplasm. The current study aimed to present a pathogenic variant in FANCA observed in three Mexican siblings, as detected by next-generation sequencing (NGS). The results of an induced chromosomal breakage test showed chromosomal breaks and radial figures, which were compatible with FA, and a normal karyotype. NGS TruSight Hereditary Cancer Panel analysis resulted in the FANCA:c.3931_3932delAG variant being classified as pathogenic according to bioinformatics analysis. The present study reports a pathogenic variant in FANCA that was found in a Mexican family with FA, in which one of the siblings exhibited a suggestive mucosa-assisted lymphoid tissue lymphoma, which is an atypical presentation of neoplasia associated with FA.

Retinal and Choroidal Circulation Impairments in Fanconi Anemia.

To evaluate the effects of Fanconi anemia (FA) on retinal and choroidal microvasculature using Optical Coherence Tomography (OCT) and Optical Coherence Tomography Angiography (OCTA). Cohort study with age-matched controls. This study included 11 eyes from 11 patients diagnosed with FA and 12 eyes from 12 age-matched healthy controls. Comprehensive ophthalmic examinations included best-corrected visual acuity (BCVA), intraocular pressure measurements, and OCT/OCTA imaging. Central macular thickness (CMT), subfoveal choroidal thickness (SCT), retinal nerve fiber layer (RNFL) thickness, and choroidal vascularity index (CVI) were measured using OCT. Superficial, deep, and choriocapillaris vessel density, foveal avascular zone (FAZ) parameters, along with radial peripapillary capillary plexus perfusion (RPCPD) and flux index (RPCFI), were assessed using OCTA. Statistical analyses were performed to compare both groups, with adjustments for axial length. Retinal and choroidal microvascular parameters, including CMT, SCT, RNFL thickness, CVI, vessel density, RPCPD, RPCFI and FAZ parameters. There was no significant difference in age or gender distribution between the FA and control groups (p = 0.98, p = 0.80, respectively). The mean age of FA patients was 19.6 ± 11.3 years (range: 6-43), while the mean age of controls was 19.4 ± 10.4 years (range: 8-40). FA patients exhibited significantly reduced SCT (252 ± 72 μm vs. 368 ± 93 μm, p=0.004) compared to controls. Vessel density in the superficial and deep capillary plexuses were significantly lower in FA patients (p=0.042 and p=0.019, respectively). Temporal (45.2 ± 2.3% vs. 48.8 ± 2.8%, p=0.047) and outer (43.6 ± 1.7% vs. 45.8 ± 1.3% p=0.037) RPCPD and temporal (0.43 ± 0.03 vs. 0.47 ± 0.02, p=0.040) and outer (0.41± 0.02 vs. 0.46± 0.02, p=0.010) RPCFI were notably reduced in FA group. CVI was comparable between groups (p=0.857), despite smaller luminal and total areas in FA patients. Fanconi anemia has a profound impact on retinal and choroidal microcirculation, characterized by reduced microvascular densities in the capillary plexi, a lower flux index in the optic nerve head, and thinning of the choroid, even in the absence of visible fundus abnormalities. These findings highlight the significant vascular effects of FA, affecting both the structural integrity and functional capacity of the retina and choroid.

Genetic buffering mechanisms in SNF2-family translocases.

SNF2-family DNA translocases, a large family of ATPases, have poorly defined roles in genomic stability. In a recent study, Feng et al. identified a synthetic lethal interaction between the SNF2 translocase SMARCAL1 and Fanconi anemia (FA) group M (FANCM), revealing a new genetic buffering mechanism that maintains genome stability by aiding DNA replication at loci enriched in simple repeats.

Q&A with Ying Wai Chan.

Ying Wai Chan spoke with Cell Reports about his journey in science and his recent paper in which he and his fellow authors identified a role of FANCM in promoting resistance to PARP inhibitors independently of the core Fanconi anemia complex.

The Significance of the Response: Beyond the Mechanics of DNA Damage and Repair-Physiological, Genetic, and Systemic Aspects of Radiosensitivity in Higher Organisms.

Classical radiation biology as we understand it clearly identifies genomic DNA as the primary target of ionizing radiation. The evidence appears rock-solid: ionizing radiation typically induces DSBs with a yield of ~30 per cell per Gy, and unrepaired DSBs are a very cytotoxic lesion. We know very well the kinetics of induction and repair of different types of DNA damage in different organisms and cell lines. And yet, higher organisms differ in their radiation sensitivity-humans can be unpredictably radiosensitive during radiotherapy; this can be due to genetic defects (e.g., ataxia telangiectasia (AT), Fanconi anemia, Nijmegen breakage syndrome (NBS), and the xeroderma pigmentosum spectrum, among others) but most often is unexplained. Among other mammals, goats (Capra hircus) appear to be very radiosensitive (LD50 = 2.4 Gy), while Mongolian gerbils (Meriones unguiculatus) are radioresistant and withstand quadruple that dose (LD50 = 10 Gy). Primary radiation lethality in mammals is due most often to hematopoietic insufficiency, which is, in the words of Dr. Theodor Fliedner, one of the pioneers of radiation hematology, "a disturbance in cellular kinetics". And yet, what makes one cell type, or one particular organism, more sensitive to ionizing radiation? The origins of radiosensitivity go above and beyond the empirical evidence and models of DNA damage and repair-as scientists, we must consider other phenomena: the radiation-induced bystander effect (RIBE), abscopal effects, and, of course, genomic instability and immunomodulation. It seems that radiosensitivity is not entirely determined by the mathematics of DNA damage and repair, and it is conceivable that radiation biology may benefit from an informed enquiry into physiology and organism-level signaling affecting radiation responses. The current article is a review of several key aspects of radiosensitivity beyond DNA damage induction and repair; it presents evidence supporting new potential venues of research for radiation biologists.

Statistical Analysis of Gene Variants for Homologous Recombination Pathways of DNA Repair leading to Cancer Susceptibility

Background: RAD51C, a critical member of the RAD51 paralog family, is essential for homologous recombination (HR)-mediated DNA repair, a pathway crucial for maintaining genomic stability. Mutations in RAD51C have been linked to cancer susceptibility, particularly in breast and ovarian cancers, where impaired DNA repair mechanisms contribute to genomic instability and tumor progression. Despite its clinical significance, the functional impact of specific RAD51C variants remains poorly understood, necessitating a comprehensive investigation into their biological implications. Methods: This study classified RAD51C gene variants into damaging and tolerant categories using computational prediction tools, including SIFT, PolyPhen, CADD, MetaLR, and Mutation Assessor. Variants were prioritized based on consensus scores and classified as high-confidence damaging variants. Correlation and agreement among tools were analyzed to refine predictions. Principal Component Analysis (PCA) and clustering methods were employed to group variants based on prediction patterns. Protein-protein interaction (PPI) networks and pathway enrichment analyses were conducted to contextualize damaging variants within broader biological systems, with a focus on their roles in HR, DNA repair, and cellular processes. Results: A total of 2526 variants were analyzed, with damaging variants showing consistent patterns across tools. Consensus scores highlighted 302 high-confidence damaging variants, which were associated with disrupted biological processes, including double-strand break repair via homologous recombination, telomere maintenance, and regulation of cell cycle checkpoints. PPI analysis revealed an interconnected network with 11 nodes and 54 edges, with a clustering coefficient of 0.982, indicating tightly coordinated interactions among DNA repair proteins. Pathway enrichment analyses identified significant associations with homologous recombination (FDR = 2.55E-17) and the Fanconi anemia pathway (FDR = 2.96E-06). Conclusion: This study provides a comprehensive framework for assessing the functional impacts of RAD51C variants by integrating computational predictions with biological analyses. The findings underscore the importance of RAD51C in HR and DNA repair pathways, offering insights into its role in genomic stability and cancer progression. These results can inform the prioritization of variants for experimental validation and guide therapeutic strategies targeting DNA repair deficiencies.

Proximity Proteomics Reveals USP44 Forms a Complex with BRCA2 in Neuroblastoma Cells and Is Required to Prevent Chromosome Breakage.

The enzyme ubiquitin-specific protease 44 (USP44) is a deubiquitinating enzyme with identified physiological roles as a tumor suppressor and an oncogene. While some binding partners and substrates are known for USP44, the identification of other interactions may improve our understanding of its role in cancer. We therefore performed a proximity biotinylation study that identified products of several known cancer genes that are associated with USP44, including a novel interaction between BRCA2 and USP44. We expressed a fusion protein that linked USP44 and mutant Escherichia coli biotin ligase BioID in SH-SY5Y neuroblastoma cells. Control experiments were performed using BioID alone. In duplicate experiments, cells were pulsed with biotin and biotinylated proteins were isolated under denaturing conditions and the proteins were identified by mass spectrometry. The resulting list of proteins were analyzed using Enrichr and cross-referenced with the COSMIC Cancer Gene Census. We validated the association with BRCA2 using immunoprecipitation. The role of USP44 in the Fanconi anemia DNA repair pathway was investigated using chromosome analysis of wild-type or Usp44-knockout cells after exposure to mitomycin C. We identified 146 proteins that were selectively retrieved by the USP44 construct and compared with cells expressing the BioID ligase alone, including 15 gene products encoded by genes on tier 1 of the COSMIC Cancer Gene Census, including BRCA2. The association between USP44 and BRCA2 was validated through immunoprecipitation. We tested the functional role of USP44 in the Fanconi anemia DNA repair pathway through chromosome breakage analysis and found that cells lacking USP44 had a significant increase in chromosome breaks and radial chromosomes. We found that high BRCA2 transcript was correlated with poor survival in neuroblastoma, likely due to its tight association with proliferation in these tumors. Our results identified novel potential binding partners and potential substrates for USP44, including several with direct roles in cancer pathogenesis. Our results identified a novel association between BRCA2 and USP44, and a previously unknown role for USP44 in the Fanconi anemia DNA repair pathway that may contribute to its role in cancer.

PRMT5-regulated splicing of DNA repair genes drives chemoresistance in breast cancer stem cells.

Breast cancer stem cells (BCSCs) are a rare cell population that is responsible for tumour initiation, metastasis and chemoresistance. Despite this, the mechanism by which BCSCs withstand genotoxic stress is largely unknown. Here, we uncover a pivotal role for the arginine methyltransferase PRMT5 in mediating BCSC chemoresistance by modulating DNA repair efficiency. Mechanistically, we identify PRMT5 as a major regulator of DNA damage response (DDR) gene splicing in BCSCs, particularly those integral to the Fanconi Anaemia and homologous recombination pathways, with PRMT5 inhibition synergising with chemotherapy to promote BCSC apoptosis. A comparison of BCSCs and their bulk cell progeny identified some shared (ATM, DDX11, EXO1, FAN1, SLX4) but many unique (ATR, RAD17, RAD51D, RUVBL1) PRMT5-dependent alternative DDR splicing events. Surprisingly, these skipped exons and retained intron events rarely lead to substantial gene expression repression, suggesting that PRMT5 inhibition predominantly results in nuclear detention of intron-containing transcripts and the production of non-canonical isoforms with compromised protein function. Since many genes within the same DDR pathway undergo deregulated splicing, this study thus reveals additional points of vulnerability and alternative combination drug strategies that could improve the therapeutic efficacy of PRMT5 inhibitors to promote BCSC eradication.

Dynamic tracking of native precursors in adult mice.

Hematopoietic dysfunction has been associated with a reduction in the number of active precursors. However, precursor quantification at homeostasis and under diseased conditions is constrained by the scarcity of available methods. To address this issue, we optimized a method for quantifying a wide range of hematopoietic precursors. Assuming the random induction of a stable label in precursors following a binomial distribution, estimates depend on the inverse correlation between precursor numbers and the variance of precursor labeling among independent samples. Experimentally validated to cover the full dynamic range of hematopoietic precursors in mice (1-105), we utilized this approach to demonstrate that thousands of precursors, which emerge after modest expansion during fetal-to-adult transition, contribute to native and perturbed hematopoiesis. We further estimated the number of precursors in a mouse model of Fanconi Anemia, showcasing how repopulation deficits can be classified as autologous (cell proliferation) and non-autologous (lack of precursor). Our results support an accessible and reliable approach for precursor quantification, emphasizing the contemporary perspective that native hematopoiesis is highly polyclonal.

Alu-Mediated Deletion of FANCA in Turkish Families With Fanconi Anemia: Evidence of a Founder Effect.

Fanconi anemia (FA) is a rare inherited bone marrow failure syndrome characterized by pancytopenia, increased susceptibility to malignancies, and a spectrum of congenital anomalies. Here, we report on eight affected individuals from six unrelated families with a large Alu-mediated intragenic deletion encompassing exons 6-31 in the FANCA gene, identified as a founder mutation in the Turkish population through haplotype analysis. This deletion, mediated by Alu repeat sequences, underscores the role of repetitive elements in FA pathogenesis. Clinical data revealed variable phenotypic presentations among affected individuals, highlighting the challenge of establishing genotype-phenotype correlations even in the presence of identical FANCA pathogenic variants. We carried out an easy and effective PCR-based diagnostic test for detecting this mutation, enabling precise diagnosis and genetic counseling for affected individuals and their families. This study provides valuable insights into the molecular mechanisms underlying FA pathogenesis and offers a practical approach for genetic diagnosis in affected individuals, particularly those of Turkish descent.

Haematopoietic gene therapy of non-conditioned patients with Fanconi anaemia-A: results from open-label phase 1/2 (FANCOLEN-1) and long-term clinical trials

Allogeneic haematopoietic stem-cell transplantation is the standard treatment for bone marrow failure (BMF) in patients with Fanconi anaemia, but transplantation-associated complications such as an increased incidence of subsequent cancer are frequent. The aim of this study was to evaluate the safety and efficacy of the infusion of autologous gene-corrected haematopoietic stem cells as an alternative therapy for these patients. This was an open-label, investigator-initiated phase 1/2 clinical trial (FANCOLEN-1) and long-term follow-up trial (up to 7 years post-treatment) in Spain. Mobilised peripheral blood (PB) CD34+ cells from nine patients with Fanconi anaemia-A in the early stages of BMF were transduced with a therapeutic FANCA-encoding lentiviral vector and re-infused without any cytotoxic conditioning treatment. The primary efficacy endpoint of FANCOLEN-1 was the engraftment of transduced cells, as defined by the detection of at least 0·1 therapeutic vector copies per nucleated cell of patient bone marrow (BM) or PB at the second year post-infusion, without this percentage having declined substantially over the previous year. The safety coprimary endpoint was adverse events during the 3 years after infusion. The completed open-label phase 1/2 and the ongoing long-term clinical trials are registered with ClinicalTrials.gov, NCT03157804; EudraCT, 2011-006100-12; and NCT04437771, respectively. There were eight evaluable treated patients with Fanconi anaemia-A. Patients were recruited between Jan 7, 2016 and April 3, 2019. The primary endpoint was met in five of the eight evaluable patients (62·50%). The median number of therapeutic vector copies per nucleated cell of patient BM and PB at the second year post-infusion was 0·18 (IQR 0·01-0·20) and 0·06 (0·01-0·19), respectively. No genotoxic events related to the gene therapy were observed. Most treatment-emergent adverse events (TEAEs) were non-serious and assessed as not related to therapeutic FANCA-encoding lentiviral vector. Nine serious adverse events (grade 3-4) were reported in six patients, one was considered related to medicinal product infusion, and all resolved without sequelae. Cytopenias and viral infections (common childhood illnesses) were the most frequently reported TEAEs. These results show for the first time that haematopoietic gene therapy without genotoxic conditioning enables sustained engraftment and reversal of BMF progression in patients with Fanconi anaemia. European Commission, Instituto de Salud Carlos III, and Rocket Pharmaceuticals.

Systematic identification of pathogenic variants of non-small cell lung cancer in the promoters of DNA-damage repair genes

Deficiency in DNA-damage repair (DDR) genes, often due to disruptive coding variants, is linked to higher cancer risk. Our previous study has revealed the association between rare loss-of-function variants in DDR genes and the risk of lung cancer. However, it is still challenging to study the predisposing role of rare regulatory variants of these genes. Based on whole-genome sequencing data from 2984 patients with non-small cell lung cancer (NSCLC) and 3020 controls, we performed massively parallel reporter assays on 1818 rare variants located in the promoters of DDR genes. Pathway- or gene-level burden analyses were performed using Firth's logistic regression or generalized linear model. We identified 750 rare functional regulatory variants (frVars) that showed allelic differences in transcriptional activity within the promoter regions of DDR genes. Interestingly, the burden of frVars was significantly elevated in cases (odds ratio [OR]=1.17, p=0.026), whereas the burden of variants prioritized solely based on bioinformatics annotation was comparable between cases and controls (OR=1.04, p=0.549). Among the frVars, 297 were down-regulated transcriptional activity (dr-frVars) and 453 were up-regulated transcriptional activity (ur-frVars); especially, dr-frVars (OR=1.30, p=0.008) rather than ur-frVars (OR=1.06, p=0.495) were significantly associated with risk of NSCLC. Individuals with NSCLC carried more dr-frVars from Fanconi anemia, homologous recombination, and nucleotide excision repair pathways. In addition, we identified seven genes (i.e., BRCA2, GTF2H1, DDB2, BLM, ALKBH2, APEX1, and RAD51B) with promoter dr-frVars that were associated with lung cancer susceptibility. Our findings indicate that functional promoter variants in DDR genes, in addition to protein-truncating variants, can be pathogenic and contribute to lung cancer susceptibility. National Natural Science Foundation of China, Youth Foundation of Jiangsu Province, Research Unit of Prospective Cohort of Cardiovascular Diseases and Cancer of Chinese Academy of Medical Sciences, and Natural Science Foundation of Jiangsu Province.

ALDH Enzymes and Hematological Diseases: A Scoping Review of Literature.

Aldehyde dehydrogenases (ALDHs) constitute a group of enzymes that catalyze the oxidation of aldehydes to carboxylic acids. The human ALDH superfamily, including 19 different isoenzymes (ALDH1A1, ALDH1A2, ALDH1A3, AHDH1B1, ALDH1L1, ALDH1L2, ALDH2, ALDH3A1, ALDH3A2, ALDH3B1, ALDH3B2, ALDH4A1, ALDH5A1, ALDH6A1, ALDH7A1, ALDH8A1, ALDH9A1, ALDHA16A1, ALDH18A1), displays different key physiological and toxicological functions, with specific tissue expression and substrate specificity. Several studies have established that ALDH are interesting markers for the identification and quantification of human hematopoietic stem cells and cancer stem cells, notably leukemic stem cells. ALDH2 is the best-documented enzyme, in this family, as having an impact on hematology, particularly myeloid malignancies. ALDH2 mainly catalyzes the detoxification of toxic aldehydes (acetaldehyde, formaldehyde). For example, ALDH2 detoxifies formaldehyde, which is produced during the differentiation of hematopoietic progenitors. The trigger of alcohol dehydrogenase 5 (also known as formaldehyde dehydrogenase or S-nitrosoglutathione reductase, ADH5/FDH/GSNOR)/ALDH2 allows to eliminate formaldehyde and ensures normal hematopoiesis. Moreover, the ALDH2*2 variant allele is the most frequent ALDH2 variant, found in 35-45% of individuals of East Asian origin. It is associated with altered acetaldehyde metabolism and is involved in several hematological diseases (aplastic anemia, bone marrow failure, myelodysplastic syndrome). This review presents current knowledge of different members of the ALDH family and their involvement in normal and malignant hematopoiesis. Focus was brought to the ALDH2 isoenzyme in congenital (Fanconi anemia, Aplastic anemia, mental retardation, and dwarfism (AMeD) syndrome, and idiopathic aplastic anemia) and acquired (acute myeloid leukemia and myelodysplastic syndrome) hematological diseases. It also describes the possibilities of using ALDH as both a biomarker and therapeutic target, to identify and eradicate leukemic stem cells in malignant diseases.

Identifying potential active ingredients from pomegranate in treating anemia: CPA3 and SOX4 are key proteins

Fanconi anemia is a rare hereditary blood disorder that usually manifests as bone marrow failure, dysplasia, cancer susceptibility and anemia. Pomegranate, as a “secret” for people in Xinjiang, China and India, is commonly used for treating different types of anemia. ObjectiveThis study aimed to identify potential proteins of FA and to discover potential drugs from pomegranates. MethodsFirstly, we downloaded gene expression datasets of myeloid cells from FA patients from the GEO database and screened for key differential genes using weighted gene co-expression network analysis and machine learning algorithms. Next, subcellular localization and external validation datasets were used to verify the reliability of genes. Finally, molecular docking and molecular dynamics simulation were used to predict potential drugs for treating FA with pomegranate. ResultsAfter screening by WGCNA and machine learning algorithms and an external validation set, SOX4 and CPA3 were retained and were highly expressed in erythroid cells of the bone marrow. Based on the above two protein structures, coumaric acid 4-O-glucoside (−8.212 Kcal/mol) and kaempferol-3-O-neohesperidoside (−7.128 Kcal/mol) were screened. ConclusionSOX4 and CPA3 can be considered as key proteins of FA, coumaric acid-4-O-glucoside and kaempferol-3-O-neohesperidin have been found to have therapeutic potency for FA.

Tuberculosis presenting with multiple osteolytic lesions in an adolescent boy with fanconi anemia

Tuberculosis (TB) is well known to present with varied clinical and radiological features. Multifocal bone involvement in tuberculosis is rare. Multiple lytic lesions raise the possibility of malignancy usually, especially, in children with predisposing conditions like Fanconi anemia. However, a thorough evaluation with repeated attempts at histopathological confirmation and microbiological evaluation is mandatory in children with unique presentations with inconclusive initial diagnostic tests. We report an adolescent boy with an underlying Fanconi anemia who presented with fever, cough and respiratory distress. On evaluation, he had pallor, wasting, features of right upper lobe consolidation and multiple lytic lesions. Initial diagnostic evaluation including bone marrow evaluation and a diagnostic biopsy from ribs and bronchoalveolar lavage failed to yield a conclusive diagnosis. After nearly 5 weeks of broad-spectrum antibiotics and antifungals, as the clinical signs were progressing, a complete re-evaluation including biopsy from the rib lesion was done and further, he was diagnosed to have skeletal tuberculosis based on the Mycobacterium Growth Indicator Tube (MGIT) culture for Acid Fast bacilli (AFB). Child was empirically started on anti-tuberculous therapy pending cultures, which have led to significant clinical improvement in 2 weeks of therapy. Low bacterial load in skeletal TB reduces the chance to diagnosis, hence making clinical judgment imperative in diagnosis; and repeated testing is mandatory when there is high index of suspicion.

Unveiling Immunotherapy Evasion in Lung Cancer: The Role of Fanconi Anemia and Stemness Genes in Shaping an Immunosuppressive Microenvironment.

The study aimed to investigate the fanconi anemia (FA)-related and stemness-related genes in lung cancer (LC) patients. Firstly, we identified stemness-related genes through weighted gene co-expression network analysis combined with TCGA database. Further combined stemness-related genes with FA-related genes to screen for prognostic-related genes. Risk score was constructed from the screened genes and comprehensive bioinformatics analyses were performed. Finally, single-cell data and in vitro experiment were used to validate our results. We screened a total of eight genes to construct a risk score. The risk score was an independent prognostic factor for LC. The validation results of multiple GEO databases were consistent with our results. Functional and pathway enrichment analysis showed that risk score was associated with cell cycle, DNA replication, DNA damage repair, and immune-related pathways. The results showed to be related to the stem cell self-renewal and proliferation. Besides, we also found that patients with higher risk scores had lower immune activity and function, and the effectiveness of immunotherapy might be poorer, with a higher rate of immune escape. Finally, our results revealed that SLC2A1 had the highest correlation with B cells in single-cell data analysis, and we validated its correlation with B cells and its expression with FA-related genes, tumor invasiveness, stemness, and drug sensitivity. Our research constructed a risk score based on FA-related and tumor stemness-related specific genes. In addition to accurately predicting the prognosis of patients with LC, the risk score may also serve as an innovative and viable predictor of immunotherapy response.