СОВРЕМЕННАЯ ТЕХНОЛОГИЯ ПОДГОТОВКИ И ПРОВЕДЕНИЯ ЛУЧЕВОЙ ТЕРАПИИ БОЛЬНЫХ ПЛОСКОКЛЕТОЧНЫМ РАКОМ АНАЛЬНОГО КАНАЛА

Squamous-cell anal cancer is a rare disease that requires a comprehensive approach in treatment and skilled professionals. Modern diagnostics is important for rational choice of treatment tactics. Radiotherapy is the cornerstone of sphincter-sparing anal cancer treatment. Radiotherapy dose, volume and duration are the key factors affecting treatment efficacy and toxicity.3D-conformal radiotherapy is a priority treatment allowing exact reproduction of treatment conditions, controlled by OBI (on-board imager) and kV X-Ray and cone-beam CT analysis. Intensity-modulated radiation therapy (IMRT) is a next-generation treatment with improved technologies, allowing better protection of normal tissues.Our experience with 21 squamous-cell anal cancer patients treated with IMRT during Nov 2011 – March 2013 is presented in this article.Эпителиальный рак анального канала является редким заболеванием, требующим в лечении комплексного подхода и квалифицированных специалистов. Для выбора рациональной комбинации лечебных методов важно использовать необходимый комплекс современных диагностических методов исследования.Основой органосохраняющего комбинированного метода лечения больных плоскоклеточным раком анального канала является лучевая терапия (ЛТ). Прогностическими факторами, влияющими на эффективность и негативные последствия комбинированного лечения, являются доза, объем и длительность проведения ЛТ.Приоритетом при проведении ЛТ является объемное 3D-планирование и использование конформной ЛТ (3D-CRT), точность воспроизведения условий ЛТ, контролируемой с помощью системы портальной визуализации в мегавольтном пучке линейного ускорителя электронов – OBI (on-board imager) и использование конического киловольтного пучка рентгеновского излучения.ЛТ с модуляцией интенсивности (IMRT) является следующей ступенью и более усовершенствованным вариантом с технологическими инновациями в конформной ЛТ, позволяющей уменьшить объем облучения нормальных тканей.В работе представлены непосредственные результаты первого опыта использования курса IMRT в комбинированном консервативном лечении 21 больного эпителиальным раком анального канала, проведенного с 11.2011 по 03.2013 в радиологическом отделении отдела радиационной онкологии ФГБУ «РОНЦ им. Н.Н. Блохина» РАМН

Efficient Differentiation of Embryonic Stem Cells into Mesodermal Precursors by BMP, Retinoic Acid and Notch Signalling

The ability to direct differentiation of mouse embryonic stem (ES) cells into specific lineages not only provides new insights into the pathways that regulate lineage selection but also has translational applications, for example in drug discovery. We set out to develop a method of differentiating ES cells into mesodermal cells at high efficiency without first having to induce embryoid body formation. ES cells were plated on a feeder layer of PA6 cells, which have membrane-associated stromal-derived inducing activity (SDIA), the molecular basis of which is currently unknown. Stimulation of ES/PA6 co-cultures with Bone Morphogenetic Protein 4 (BMP4) both favoured self-renewal of ES cells and induced differentiation into a Desmin and Nestin double positive cell population. Combined stimulation with BMP4 and all-trans Retinoic Acid (RA) inhibited self-renewal and resulted in 90% of cells expressing Desmin and Nestin. Quantitative reverse transcription-polymerase chain reaction (qPCR) analysis confirmed that the cells were of mesodermal origin and expressed markers of mesenchymal and smooth muscle cells. BMP4 activation of a MAD-homolog (Smad)-dependent reporter in undifferentiated ES cells was attenuated by co-stimulation with RA and co-culture with PA6 cells. The Notch ligand Jag1 was expressed in PA6 cells and inhibition of Notch signalling blocked the differentiation inducing activity of PA6 cells. Our data suggest that mesodermal differentiation is regulated by the level of Smad activity as a result of inputs from BMP4, RA and the Notch pathway

Specific Marking of hESCs-Derived Hematopoietic Lineage by WAS-Promoter Driven Lentiviral Vectors

Genetic manipulation of human embryonic stem cells (hESCs) is instrumental for tracing lineage commitment and to studying human development. Here we used hematopoietic-specific Wiskott-Aldrich syndrome gene (WAS)-promoter driven lentiviral vectors (LVs) to achieve highly specific gene expression in hESCs-derived hematopoietic cells. We first demonstrated that endogenous WAS gene was not expressed in undifferentiated hESCs but was evident in hemogenic progenitors (CD45−CD31+CD34+) and hematopoietic cells (CD45+). Accordingly, WAS-promoter driven LVs were unable to express the eGFP transgene in undifferentiated hESCs. eGFP+ cells only appeared after embryoid body (EB) hematopoietic differentiation. The phenotypic analysis of the eGFP+ cells showed marking of different subpopulations at different days of differentiation. At days 10–15, AWE LVs tag hemogenic and hematopoietic progenitors cells (CD45−CD31+CD34dim and CD45+CD31+CD34dim) emerging from hESCs and at day 22 its expression became restricted to mature hematopoietic cells (CD45+CD33+). Surprisingly, at day 10 of differentiation, the AWE vector also marked CD45−CD31low/−CD34− cells, a population that disappeared at later stages of differentiation. We showed that the eGFP+CD45−CD31+ population generate 5 times more CD45+ cells than the eGFP−CD45−CD31+ indicating that the AWE vector was identifying a subpopulation inside the CD45−CD31+ cells with higher hemogenic capacity. We also showed generation of CD45+ cells from the eGFP+CD45−CD31low/−CD34− population but not from the eGFP−CD45−CD31low/−CD34− cells. This is, to our knowledge, the first report of a gene transfer vector which specifically labels hemogenic progenitors and hematopoietic cells emerging from hESCs. We propose the use of WAS-promoter driven LVs as a novel tool to studying human hematopoietic development

MODERN TECHNOLOGIES IN SQUAMOUS-CELL ANAL CANCER RADIOTHERAPY PLANNING AND TREATMENT

Squamous-cell anal cancer is a rare disease that requires a comprehensive approach in treatment and skilled professionals. Modern diagnostics is important for rational choice of treatment tactics. Radiotherapy is the cornerstone of sphincter-sparing anal cancer treatment. Radiotherapy dose, volume and duration are the key factors affecting treatment efficacy and toxicity.3D-conformal radiotherapy is a priority treatment allowing exact reproduction of treatment conditions, controlled by OBI (on-board imager) and kV X-Ray and cone-beam CT analysis. Intensity-modulated radiation therapy (IMRT) is a next-generation treatment with improved technologies, allowing better protection of normal tissues.Our experience with 21 squamous-cell anal cancer patients treated with IMRT during Nov 2011 – March 2013 is presented in this article

Preliminary results of accelerated partial breast irradiation in the combination organ-saving treatment of patients with early-stage breast cancer

Objective: to develop and introduce an accelerated partial breast irradiation procedure in patients with early-stage breast cancer after organ-saving operations using three-dimensional conformal teleradiotherapy (3D CRT) and to assess the preliminary results of treat- ment.Subjects and methods: the trial enrolled 48 patients with verified T1-2N0-1micM0 stage breast cancer who received organ-saving treatment in the N.N. Blokhin Russian Cancer Research Center, Russian Academy of Medical Sciences, in 2008–2012. The median age of the patients was 63 years. All the patients underwent organ-saving operations with subsequent 3D CRT applied to the resected tumor bed with margins. Radiotherapy with a dose of 2.5 Gy was performed twice daily; the daily dose was 5 Gy; the course dose was 40 Gy. Indications for accelerated partial irradiation of the tumor bed were listed; the data of randomized trials were given.Results. The median follow-up was 27 months; locoregional recurrences or distant metastases were found in no patients. Late radia- tion skin and soft tissue injuries were seen in no cases. All the patients were ascertained to have early-stage grade I radiation cutane- ous injuries according to the RTOG-EORTC classification. Excellent, good, fair, and poor cosmetic results were obtained in 12 (25 %), 34 (71 %), 2 (4 %), and 0 patients, respectively.Conclusion. The developed accelerated partial breast irradiation procedure in patients with early-stage breast cancer after organ- saving operations at the median follow-up of 27 months showed good results, such as no local recurrences or distant metastases. Excel- lent and good cosmetic results were noted in 96 % of the patients