Nanoemulsions as Gene Delivery in Mucopolysaccharidosis Type I-A Mini-Review.

Mucopolysaccharidosis type I (MPS I) is a rare monogenic disease in which glycosaminoglycans' abnormal metabolism leads to the storage of heparan sulfate and dermatan sulfate in various tissues. It causes its damage and impairment. Patients with the severe form of MPS I usually do not live up to the age of ten. Currently, the therapy is based on multidisciplinary care and enzyme replacement therapy or hematopoietic stem cell transplantation. Applying gene therapy might benefit the MPS I patients because it overcomes the typical limitations of standard treatments. Nanoparticles, including nanoemulsions, are used more and more in medicine to deliver a particular drug to the target cells. It allows for creating a specific, efficient therapy method in MPS I and other lysosomal storage disorders. This article briefly presents the basics of nanoemulsions and discusses the current state of knowledge about their usage in mucopolysaccharidosis type I.

Exome Sequencing Reveals Novel Variants and Expands the Genetic Landscape for Congenital Microcephaly.

Congenital microcephaly causes smaller than average head circumference relative to age, sex and ethnicity and is most usually associated with a variety of neurodevelopmental disorders. The underlying etiology is highly heterogeneous and can be either environmental or genetic. Disruption of any one of multiple biological processes, such as those underlying neurogenesis, cell cycle and division, DNA repair or transcription regulation, can result in microcephaly. This etiological heterogeneity manifests in a clinical variability and presents a major diagnostic and therapeutic challenge, leaving an unacceptably large proportion of over half of microcephaly patients without molecular diagnosis. To elucidate the clinical and genetic landscapes of congenital microcephaly, we sequenced the exomes of 191 clinically diagnosed patients with microcephaly as one of the features. We established a molecular basis for microcephaly in 71 patients (37%), and detected novel variants in five high confidence candidate genes previously unassociated with this condition. We report a large number of patients with mutations in tubulin-related genes in our cohort as well as higher incidence of pathogenic mutations in MCPH genes. Our study expands the phenotypic and genetic landscape of microcephaly, facilitating differential clinical diagnoses for disorders associated with most commonly disrupted genes in our cohort.

Gene Therapy for Mucopolysaccharidosis Type II-A Review of the Current Possibilities.

Mucopolysaccharidosis type II (MPS II) is a lysosomal storage disorder based on a mutation in the IDS gene that encodes iduronate 2-sulphatase. As a result, there is an accumulation of glycosaminoglycans-heparan sulphate and dermatan sulphate-in almost all body tissues, which leads to their dysfunction. Currently, the primary treatment is enzyme replacement therapy, which improves the course of the disease by reducing somatic symptoms, including hepatomegaly and splenomegaly. The enzyme, however, does not cross the blood-brain barrier, and no improvement in the function of the central nervous system has been observed in patients with the severe form of the disease. An alternative method of treatment that solves typical problems of enzyme replacement therapy is gene therapy, i.e., delivery of the correct gene to target cells through an appropriate vector. Much progress has been made in applying gene therapy for MPS II, from cellular models to human clinical trials. In this article, we briefly present the history and basics of gene therapy and discuss the current state of knowledge about the methods of this therapy in mucopolysaccharidosis type II.

Coincidence of 3-methylglutaconic aciduria and duplication 5q - a case report and literature review.

3-methylglutaconic aciduria includes a heterogeneous group of inborn errors of metabolism. The disease may have various clinical presentations, as can duplication 5q. We present the case of a 13-year-old boy with 3-methylglutaconic aciduria and duplication 5q. The main symptoms included myopathy, weakness, spastic paresis intensified mostly in the lower limbs, and intellectual disability. Additional studies showed elevated levels of 3-methylglutaconic acid in urine and ammonia in plasma. A duplication in region 5q23.3q31.1 was found in array-based comparative genomic hybridization. Next-generation sequencing did not reveal any pathological mutation. On the basis of the clinical picture and the results of biochemical and genetic tests 3-methylglutaconic aciduria type IV with duplication 5q was diagnosed.

A Dual Face of APE1 in the Maintenance of Genetic Stability in Monocytes: An Overview of the Current Status and Future Perspectives.

Monocytes, which play a crucial role in the immune system, are characterized by an enormous sensitivity to oxidative stress. As they lack four key proteins responsible for DNA damage response (DDR) pathways, they are especially prone to reactive oxygen species (ROS) exposure leading to oxidative DNA lesions and, consequently, ROS-driven apoptosis. Although such a phenomenon is of important biological significance in the regulation of monocyte/macrophage/dendritic cells' balance, it also a challenge for monocytic mechanisms that have to provide and maintain genetic stability of its own DNA. Interestingly, apurinic/apyrimidinic endonuclease 1 (APE1), which is one of the key proteins in two DDR mechanisms, base excision repair (BER) and non-homologous end joining (NHEJ) pathways, operates in monocytic cells, although both BER and NHEJ are impaired in these cells. Thus, on the one hand, APE1 endonucleolytic activity leads to enhanced levels of both single- and double-strand DNA breaks (SSDs and DSBs, respectively) in monocytic DNA that remain unrepaired because of the impaired BER and NHEJ. On the other hand, there is some experimental evidence suggesting that APE1 is a crucial player in monocytic genome maintenance and stability through different molecular mechanisms, including induction of cytoprotective and antioxidant genes. Here, the dual face of APE1 is discussed.

RNF6 as an Oncogene and Potential Therapeutic Target-A Review.

The RNF6 gene encodes Ring Finger Protein 6 (RNF6), which functions as a ubiquitin ligase. Its functions are not entirely known, but research shows that it is involved in human cancer development. Initially, this gene was considered to be a tumor suppressor. Numerous statistical analyses on cell lines and animals indicate, however, that RNF6 functions as an oncogene, involved in signaling pathways, including SHP1/STAT3, AKT/mTOR, Wnt/ß-catenin, or ERα/Bcl-xL. Due to this fact, it has become a potential prognostic factor and therapeutic target. Studies in tumor cells and model organisms using inhibitors such as total saponins from Paris forrestii (TSPf), ellagic acid, or microRNA molecules show the effectiveness of inhibiting RNF6, and through it, the pathways of tumor cell proliferation. The results of the currently available studies are promising, but the function of RNF6 is not fully understood. More research is needed to assess the role of RNF6 and to check the safety and efficacy of inhibitors.

Comprehensive genomic analysis of patients with disorders of cerebral cortical development.

Malformations of cortical development (MCDs) manifest with structural brain anomalies that lead to neurologic sequelae, including epilepsy, cerebral palsy, developmental delay, and intellectual disability. To investigate the underlying genetic architecture of patients with disorders of cerebral cortical development, a cohort of 54 patients demonstrating neuroradiologic signs of MCDs was investigated. Individual genomes were interrogated for single-nucleotide variants (SNV) and copy number variants (CNV) with whole-exome sequencing and chromosomal microarray studies. Variation affecting known MCDs-associated genes was found in 16/54 cases, including 11 patients with SNV, 2 patients with CNV, and 3 patients with both CNV and SNV, at distinct loci. Diagnostic pathogenic SNV and potentially damaging variants of unknown significance (VUS) were identified in two groups of seven individuals each. We demonstrated that de novo variants are important among patients with MCDs as they were identified in 10/16 individuals with a molecular diagnosis. Three patients showed changes in known MCDs genes  and a clinical phenotype beyond the usual characteristics observed, i.e., phenotypic expansion, for a particular known disease gene clinical entity. We also discovered 2 likely candidate genes, CDH4, and ASTN1, with human and animal studies supporting their roles in brain development, and 5 potential candidate genes. Our findings emphasize genetic heterogeneity of MCDs disorders and postulate potential novel candidate genes involved in cerebral cortical development.

Variable expressivity of the phenotype in two families with brachydactyly type E, craniofacial dysmorphism, short stature and delayed bone age caused by novel heterozygous mutations in the PTHLH gene.

Brachydactyly refers to shortening of digits due to hypoplasia or aplasia of bones forming the hands and/or feet. Isolated brachydactyly type E (BDE), which is characterized by shortened metacarpals and/or metatarsals, results in a small proportion of patients from HOXD13 or PTHLH mutations, although in the majority of cases molecular lesion remains unknown. BDE, like other brachydactylies, shows clinical heterogeneity with highly variable intrafamilial and interindividual expressivity. In this study, we investigated two Polish cases (one familial and one sporadic) presenting with BDE and additional symptoms due to novel PTHLH mutations. Apart from BDE, the affected family showed short stature, mild craniofacial dysmorphism and delayed bone age. Sanger sequencing of PTHLH revealed a novel heterozygous frameshift mutation c.258delC(p.N87Tfs*18) in two affected individuals and one relative manifesting mild brachydactyly. The sporadic patient, in addition to BDE, presented with craniofacial dysmorphism, normal stature and bone age, and was demonstrated to carry a de novo heterozygous c.166C>T(p.R56*) mutation. Our paper reports on the two novel truncating PTHLH variants, resulting in variable combination of BDE and other symptoms. Data shown here expand the knowledge on the phenotypic presentation of PTHLH mutations, highlighting significant clinical variability and incomplete penetrance of the PTHLH-related symptoms.

Apolipoprotein E genotype and LRP1 polymorphisms in patients with different clinical types of metachromatic leukodystrophy.

Metachromatic leukodystrophy (MLD) is a severe, neurodegenerative, metabolic disease which is caused by deficient activity of arylsulfatase A (ARSA). Sulfatides and other substrates of ARSA are stored in central and peripheral nervous systems, and in some other organs. Accumulated sulfatides are especially toxic to oligodendrocytes and Schwann cells leading to progressive demyelination. The kind of apolipoprotein E (apoE) isoform is of essential significance for the modulation of sulfatide quantity in the brain as apoE4 contains more sulfatides than apoE3. Taking into consideration the fact that apoE4 leads to the loss of sulfatides in CSF of Alzheimer's disease patients, we examined if apoE isoforms display any impact on clinical outcome in patients with different forms of MLD in whom sulfatides accumulate. The significant association of age at the onset of MLD symptoms with APOE ε2/ε3/ε4 and LRP1 c.766C>T polymorphisms was shown in multivariate stepwise regression analysis, in which other factors known to affect age at onset of the disease, i.e. clinical type of MLD, family connection of the patient and sex were also analyzed. As expected, the clinical type of MLD explained about 80% of the variance of the dependent variable. The impact of both polymorphisms on age of onset of the disease was considerably lower: 2.0% in the case of APOE polymorphism and 1.0% in the case of LRP1 polymorphism. Thus, the clinical outcome in MLD patients is related principally to the genotype of the ARSA gene, while the polymorphisms in the APOE and LRP1 genes are only slightly modifying factors.

MURCS Association with Partial Duplication of the Distal Long Chromosome 5 and Unilateral Ovarian Agenesis.

A combination of the congenital abnormalities, Müllerian duct aplasia, renal aplasia, and cervicothoracic somite dysplasia, is defined as the MURCS association. Various genetic defects have been described in the MURCS association so far, yet the unambiguous molecular basis of these disorders has not been established. We report the case of an 18-year-old woman who presented with primary amenorrhea, right kidney, Arnold-Chiari malformation, and Klippel-Feil syndrome. In addition, the patient showed the following unusual features: right ovarian and Skenes gland agenesis, cubitus valgus with hyperextension and decreased range of motion at elbows, and facial changes. Moreover, the performed DNA analysis showed interstitial duplication in chromosome 5 (5q35.1). In the duplicated region, there are genes whose function is not well known. It is thought that they have an influence on the early stages of development and their joining in the later period can lead to neoplastic disorders, especially leukemias.

[Selected problems of the diagnosis of Turner's syndrome]. / Wybrane aspekty rozpoznania zespolu Turnera.

UNLABELLED: Turner's syndrome (TS) can be diagnosed in one out of 2130 females. The aim of the current study was to collect data about circumstances associated with the diagnosis of the syndrome. The material consisted of 177 female patients from the whole Polish territory, that turned to the out-patient Clinic for Women with Turner's syndrome, Specialist Hospital No 2 Bytom. Patients underwent physical examination and all relevant clinical information were analysed. The mean age when the abnormal symptoms were found was 8.5 years. Only 50% of those, who first turned their attention to these symptoms, were the health care workers. The initial symptoms were quite variable. Classical symptoms, i.e. short stature and amenorrhea were present respectively in only 41% and 8% of patients. In 16% of patients the diagnosis was significantly delayed, because, due to a lack of symptoms TS was not suspected. The latest cases were diagnosed at the age of 14.6 +/- 8.2 years. When it came to analysing karyotypes, simple chromosome X monosomy was present in 56% of patients and in 24% of cases mosaic karyotypes were found. Studies were performed in 14 cities in Poland. CONCLUSIONS: 1. In most cases diagnosis of TS was markedly delayed. 2. More efforts are required in order to perform karyotype studies in younger patients then it is now. Also molecular studies should become more easily available, since they are a necessary supplement to cytogenetic studies.

Quantitative ultrasound and peripheral bone densitometry in patients with genetic disorders.

The aim of the study was comparison of quantitative ultrasound and densitometric peripheral measurements in subjects with genetic disorders. The study included 52 subjects (35 boys and 17 girls) in mean age 13.1 +/- 4.8 y. Patients with following disorders were evaluated: Down syndrome (n = 21), Martin-Bell syndrome (n = 14) and other (n = 17). There were no additional factors potentially influencing bone metabolism. Bone status was assessed by quantitative ultrasound at the hand phalanges using DBM Sonic 1200 (IGEA, Italy), which measures amplitude-dependent speed of sound (Ad-SoS [m/s]) and bone densitometry at the calcaneus and forearm by the use of PIXI (GE, USA), which measures bone mineral density (BMD, g/cm2). Ad-SoS correlated significantly with forearm and calcaneus BMD in the whole group (r = 0.66, p < 0.000001 and r = 0.51, p < 0.0001, respectively), in females (r = 0.58, p < 0.05 and r = 0.5, p < 0.05) and in males (r = 0.70, p < 0.000001 and r = 0.54, p < 0.001). Calcaneus BMD correlated with wrist BMD in the whole group, in females and males: r = 0.66, p < 0.000001, r = 0.67, p < 0.01 and r = 0.75, p < 0.0001, respectively. These coefficients of correlation were compared and did not reveal significant differences in the whole group and in the gender subgroups. ROC analysis of Ad-SoS values versus calcaneus and forearm BMD showed area under curve 0.89 for forearm BMD and 0.79 for calcaneus BMD (subjects with Ad-SoS T-score below -3.2 were considered as abnormal). Age correlated significantly with Ad-SoS, forearm and calcaneus BMD (r ranged from 0.53 to 0.9, p from 0.05 to 0.000001). In all patients and males, age more strongly influenced Ad-SoS than calcaneus BMD (p < 0.05). Also, body weight and height correlated significantly with Ad-SoS, forearm and calcaneus BMD, except for correlation between forearm BMD and height in female patients (r from 0.58 to 0.84, p < 0.05). Generally, in multiple stepwise regression analysis of age and body size on skeletal parameters, age had positive influence, and body size was a positive or a negative factor. In conclusion, both quantitative ultrasound and peripheral densitometry may be recommended for the assessment of skeletal status in subjects with genetic disorders, although measurements of phalanges seem to be more sensitive for detecting age-related bone changes.

Longitudinal changes in ultrasound measurements: a parallel study in subjects with genetic disorders and healthy controls.

Disturbances in skeletal status in subjects with genetic disorder may increase their fracture risk. The aim of the study was longitudinal observation of phalangeal speed of sound changes across the bone over a period of 2 y in 24 patients (14 boys and 10 girls, mean age 9.63 +/- 1.8 y.) and 24 age-matched healthy controls (14 boys and 10 girls, mean age 9.65 +/- 1.71 y.). Weight and height did not differ between patients and controls at baseline and follow-up. Patients with the following disorders were evaluated: 7 with Down syndrome, 6 nonspecific mental retardations of unknown etiology, 5 Martin-Bell syndrome and 6 with other diseases. In patients and controls, no factors potentially influencing bone metabolism (except for genetic disorder) were present. Bone status was assessed by quantitative ultrasound at hand phalanges using DBM Sonic 1200 (IGEA, Carpi, Italy), which measures amplitude-dependent speed of sound (Ad-SoS [m/s]). At baseline, Ad-SoS and Z-score were significantly lower in patients than in controls (1892 +/- 51 m/s versus 1936 +/- 43 m/s, p < 0.01 and -1.47 +/- 1.43 versus -0.14 +/- 1.04, p < 0.001, respectively. In follow-up, Ad-SoS and Z-score increased significantly in patients (1892 +/- 51 m/s to 1934 +/- 48 m/s, p < 0.0001 and -1.47 +/- 1.43 to -0.76 +/- 1.00, p < 0.01, respectively) and in controls (1936 +/- 43 m/s to 1976 +/- 60 m/s and -0.14 +/- 1.04 to 0.31 +/- 1.08, p < 0.05, respectively). Follow-up Ad-SoS and Z-Score were significantly lower in patients (p < 0.01). Longitudinal changes in Ad-SoS, Z-score weight did not differ between patients and control, and height increased more in controls (13.2 +/- 2.8 cm versus 11.4 +/- 5.9 cm, p < 0.05)+). In patients, Ad-SoS increased by 42 m/s (2.22%), and in controls increased by 40 m/s (2.07%). Difference in Ad-SoS between patients and controls was 44 m/s at baseline and 42 m/s at follow-up. Using the value of the least significant change (LSC = 20.5 m/s), in 16 patients (67%) and in 18 controls (75%) Ad-SoS showed an increase greater than the LSC, in one control (4%) a decrease greater than the LSC and in rest of subjects studied remained unchanged (33% patients and 19% controls) over a period of observation. In conclusion, despite comparable improvement in measured ultrasound parameter in patients and controls observed over a study duration, the difference between them remained stable.

[Fetal warfarin syndrome in twin pregnancy]. / Plodowy zespól warfarynowy w przebiegu ciazy blizniaczej.

We describe a pair of twins, whose mother was being treated by oral anticoagulant drugs, as a result of having received mitral heart valve implantation in the past. The male twins monochorionic, monoamniotic--but one infant showed the features of fetal warfarin syndrome. In the study we discussed the pharmacogentetics and individual variation in the human metabolism during treatment with warfarin-perinatal growth and prevalence of congenital malformations. We analysed the threats to the fetus and mother, connected with administration of anticoagulant drugs.

[Partial lipodystrophy with C3 complement deficiency in 8 years old girl]. / Zespól lipodystrofii ograniczonej - czesciowej, z niedoborem skladowej c3 dopelniacza u 8-letniej dziewczynki.

Lipodystrophy is a rare, heterogenic disorder, leading to the loss of adipose tissue. Several main types of the disease are distinguished according to age of onset and localization of fat atrophy. The authors present the case of 8 years old girl with partial lipodystrophy, C3 complement deficiency and autoimmunologic disorder.

Schimke immuno-osseous dysplasia: two cases.

We report two patients with Schimke immuno-osseous dysplasia (SIOD). SIOD is characterised by growth retardation, renal failure, spondylo-epiphyseal dysplasia, specific phenotype and defective cellular immunity. These two children demonstrated a bone dysplasia with characteristic radiographic appearances. We postulate that SIOD should be considered in all cases of growth failure with an unclassifiable bone dysplasia. Repeated urine tests for proteinuria could be helpful in reaching the correct diagnosis.