Gene Therapy Medicinal Products: Non-clinical Safety Studies

Currently, gene therapy medicinal products (GTMPs) are actively developed in many countries, including the Russian Federation. However, the use of GTMPs raises class-specific safety concerns.The aim of the study was to determine the main requirements for non-clinical safety testing of GTMPs, to identify risks associated with these medicinal products, to establish criteria for expert assessments, and to find optimisation opportunities for GTMP non-clinical safety programmes, using Russian and international experience in the assessment of submissions and the registration of medicinal products of this class.The Russian Federation, the Eurasian Economic Union, the European Union, and the United States have created regulatory frameworks governing the lifecycle of GTMPs and continue improving these frameworks. The properties of GTMPs may create unique safety issues, such as insertional mutagenesis, unregulated transgene expression, long-term persistence and off-target spread, vertical germline transmission, and environmental risks. To account for these issues, a comprehensive non-clinical safety programme for GTMPs may require additional special studies along with the standard ones. This review focuses on the main approaches to designing non-cellular GTMP safety studies and evaluating the obtained results. The authors identified improvement opportunities for and problematic aspects of study design, as well as conditions for and limitations of non-clinical data extrapolation and clinical safety profile prediction. The continuous improvement and updating of the regulatory frameworks governing non-clinical studies of GTMPs mean that developers of non-clinical safety programmes for GTMPs should use all their experience, as well as relevant national and international guidelines and recommendations

Study of the relationship between professional self-determination and communication skills in adolescents and young men

The relevance of the research is based on the need to study the factors affecting the professional self-determination of adolescents and young men, including their communicative skills and the characteristics of peer groups, to which young people belong and are affected by. The aim of the research was to study the interrelationship between the indicators of professional self-determination and the communication skill levels in adolescents and young men. Hypothesis of the research: 1. In adolescent boys, unlike teenage girls, the choice of profession is more uncertain. 2. Schoolchildren and students have significant links between the indicators of professional self-determination and their communication skill levels. 3. There is a link between the indicators of professional self-determination and belonging to the group. In the empirical study, the following methods were used: "The Personal Professional Path" questionnaire edited by N. Pryazhnikov, the "Differential-diagnostic Questionnaire" by E. Klimova, the method "Communication and Organizational Propensity" (COS-2) by V. Sinyavsky and B. Fedorishin, the "Sociometry" Method by J. Moreno. The sample of 159 people consisted of students of the 7th and 10th grades, students of the Ural Federal University. The external and internal motives of choosing a profession by adolescents and young men were examined and empirically studied, secondly, the relationship between the choice of profession and professional inclinations, and thirdly, the interrelation of communicative and professional inclinations. The first hypothesis was confirmed by the positive correlation between communicative inclinations and professional intentions in the category "man-man" in respondents. The second hypothesis was partially confirmed: in two classes and in one student group, the differences between the mini-groups were determined by the criterion of professional inclinations, i.e. professional inclinations can be a factor uniting the students in mini-groups, but the final professional choice within the groups is different, therefore, determined by other factors.Актуальность исследования связана с необходимостью изучения факторов, влияющих на профессиональное самоопределение подростков и юношей, к числу которых относятся их коммуникативное качества, а также особенности групп сверстников, к которым принадлежат молодые люди и испытывают на себе их влияние. Цель исследования заключалась в изучении взаимосвязи показателей профессионального самоопределения и сформированности коммуникативных качеств личности в подростковом и юношеском возрасте. Гипотезы исследования: 1. У подростков-юношей, в отличие от подростков-девушек, выбор профессии отличается большей неопределенностью. 2. У школьников и студентов имеются значимые связи между показателями профессионального самоопределения и сформированностью коммуникативных качеств. 3. Существует связь между показателями профессионального самоопределения и принадлежностью к группе. В эмпирическом исследовании использовались следующие методики: анкета «Личный профессиональный путь» под редакцией Н. С. Пряжникова, «Дифференциально-диагностический опросник» Е. А. Климова, методика «Коммуникативные и организационные склонности» (КОС – 2) В. В. Синявский, Б. А. Федоришина, методика «Социометрия» Дж. Морено. Выборка в количестве 159 человек состояла из учащихся 7-х и 10-х классов, студентов Уральского федерального университета. Были рассмотрены и эмпирически исследованы внешние и внутренние мотивы выбора профессии у подростков и юношей, во-вторых, взаимосвязь выбора профессии и профессиональных склонностей, в-третьих, взаимосвязь коммуникативных и профессиональных склонностей. Первая гипотеза подтверждена наличием у респондентов положительной корреляции между коммуникативными склонностями и профессиональными намерениями в категории «человек – человек». Вторая гипотеза подтвердилась частично: в двух классах и одной студенческой группе выявлены различия между мини-группами по критерию профессиональных склонностей, т.е. профессиональные склонности могут быть фактором, объединяющим в мини-группы учащихся, но окончательный профессиональный выбор внутри групп различен, следовательно, определяется и другими факторами

Human Secreted Ly-6/uPAR Related Protein-1 (SLURP-1) Is a Selective Allosteric Antagonist of α7 Nicotinic Acetylcholine Receptor

SLURP-1 is a secreted toxin-like Ly-6/uPAR protein found in epithelium, sensory neurons and immune cells. Point mutations in the slurp-1 gene cause the autosomal inflammation skin disease Mal de Meleda. SLURP-1 is considered an autocrine/paracrine hormone that regulates growth and differentiation of keratinocytes and controls inflammation and malignant cell transformation. The majority of previous studies of SLURP-1 have been made using fusion constructs containing, in addition to the native protein, extra polypeptide sequences. Here we describe the activity and pharmacological profile of a recombinant analogue of human SLURP-1 (rSLURP-1) differing from the native protein only by one additional N-terminal Met residue. rSLURP-1 significantly inhibited proliferation (up to ~ 40%, EC50 ~ 4 nM) of human oral keratinocytes (Het-1A cells). Application of mecamylamine and atropine,--non-selective inhibitors of nicotinic acetylcholine receptors (nAChRs) and muscarinic acetylcholine receptors, respectively, and anti-α7-nAChRs antibodies revealed α7 type nAChRs as an rSLURP-1 target in keratinocytes. Using affinity purification from human cortical extracts, we confirmed that rSLURP-1 binds selectively to the α7-nAChRs. Exposure of Xenopus oocytes expressing α7-nAChRs to rSLURP-1 caused a significant non-competitive inhibition of the response to acetylcholine (up to ~ 70%, IC50 ~ 1 μM). It was shown that rSLURP-1 binds to α7-nAChRs overexpressed in GH4Cl cells, but does not compete with 125I-α-bungarotoxin for binding to the receptor. These findings imply an allosteric antagonist-like mode of SLURP-1 interaction with α7-nAChRs outside the classical ligand-binding site. Contrary to rSLURP-1, other inhibitors of α7-nAChRs (mecamylamine, α-bungarotoxin and Lynx1) did not suppress the proliferation of keratinocytes. Moreover, the co-application of α-bungarotoxin with rSLURP-1 did not influence antiproliferative activity of the latter. This supports the hypothesis that the antiproliferative activity of SLURP-1 is related to 'metabotropic' signaling pathway through α7-nAChR, that activates intracellular signaling cascades without opening the receptor channel

Resistance to thyroid hormone is modulated in vivo by the nuclear receptor corepressor (NCOR1)

Mutations in the ligand-binding domain of the thyroid hormone receptor β (TRβ) lead to resistance to thyroid hormone (RTH). These TRβ mutants function in a dominant-negative fashion to interfere with the transcription activity of wild-type thyroid hormone receptors (TRs), leading to dysregulation of the pituitary–thyroid axis and resistance in peripheral tissues. The molecular mechanism by which TRβ mutants cause RTH has been postulated to be an inability of the mutants to properly release the nuclear corepressors (NCORs), thereby inhibiting thyroid hormone (TH)-mediated transcription activity. To test this hypothesis in vivo, we crossed ThrbPV mice (a model of RTH) expressing a human TRβ mutant (PV) with mice expressing a mutant Ncor1 allele (Ncor1ΔID mice) that cannot recruit a TR or a PV mutant. Remarkably, in the presence of NCOR1ΔID, the abnormally elevated thyroid-stimulating hormone and TH levels found in ThrbPV mice were modestly but significantly corrected. Furthermore, thyroid hyperplasia, weight loss, and other hallmarks of RTH were also partially reverted in mice expressing NCOR1ΔID. Taken together, these data suggest that the aberrant recruitment of NCOR1 by RTH TRβ mutants leads to clinical RTH in humans. The present study suggests that therapies aimed at the TR–NCOR1 interaction or its downstream actions could be tested as potential targets in treating RTH

rSLURP-1 binds α7 nAChR subunits in the human brain.

<p>Affinity purification was performed using magnetic beads covalently coupled with rSLURP-1 or non-coupled beads (Ctrl) on human temporal cortical homogenates (n = 2). Samples were submitted to gel electrophoresis and Western blotting along with samples of the homogenate used for affinity purification (Input) and the remaining homogenate after affinity purification (Output) followed by detection of nAChR subunits.</p