Prof. Iana Tsoneva, Institute of Prof. Iana Tsoneva, Institute of Biophysics and Biomedical Biophysics and Biomedical

Engineering, Bulg. Acad. Sci., “Acad. Engineering, Bulg. Acad. Sci., “Acad. G. Bonchev” Str., bl. 21, 1113 Sofia, G. Bonchev” Str., bl. 21, 1113 Sofia,

Bulgaria, Bulgaria, MC member of COST MC member of COST Action TDAction TD11041104;;

Our results from participation in Our results from participation in European network for development European network for development

of electroof electroррorationoration--base technologiesbase technologies ((EPEP44BioBio22MedMed) 2012-2016) 2012-2016, , Chair of Chair of

the Actionthe Action – – ProfProf.. Damijan MIKLAVCIC Damijan MIKLAVCIC (SI) (SI)

TheThe kick kick off meeting off meeting was on was on –t–thehe April 9, 201 April 9, 2012, but the first financial 2, but the first financial

year starts on the 1-st July year starts on the 1-st July

20120122..

The project The project brinbrinss together 445 together 445 participants from 150 institutions participants from 150 institutions

from around the from around the world (32 countries) by now.world (32 countries) by now.

In the beginning the countriesIn the beginning the countries were 22were 22

1.To bring together and 1.To bring together and coordinate Europeancoordinate European community community researchresearch in in different domains of electroporation different domains of electroporation 2.2.To promote existing and To promote existing and establish new links with industrial establish new links with industrial partnerspartners and and synchronisesynchronise academic and academic and industrial research industrial research 3.3.To To optimise and further develop existing electroporation-based optimise and further develop existing electroporation-based applications and explore new applicationsapplications and explore new applications in in biology, medicine, biology, medicine, pharmacy and food industrypharmacy and food industry3.3.To provide comprehensive To provide comprehensive training for studentstraining for students4.4.To To stimulate and promote collaborative researchstimulate and promote collaborative research by structuring and by structuring and developing proposals for futuredeveloping proposals for future applications of additional funding on the applications of additional funding on the national and European levelnational and European level5.5.To synthesize and To synthesize and disseminate new and updated kdisseminate new and updated knonowledgewledge on on mechanisms of electroporation, mechanisms of electroporation, newnew applications and technological applications and technological developmentsdevelopments

EP4Bio2Med Objectives(from Miclavcic poster)

WG of COST TD1104 in paper of D. Miclavcic, in J Membrane Biol (2012) 245:591–598, DOI 10.1007/s00232-012-9493-8.

Gender balance: 34% group leaders are female researchers Statistics is given by D. Miclavcic

www.electroporation.netwww.electroporation.netNewsletter No. 17 Newsletter No. 17 , October 2013, October 2013

During the last year, there were 8,379During the last year, there were 8,379visits to the site, made by 3,793visits to the site, made by 3,793 unique unique

visitors. visitors.

According Prof. According Prof. Roland Pochet,Roland Pochet,who is also BMBS who is also BMBS ((Biomedicine and Molecular Biomedicine and Molecular

BiosciencesBiosciences) ) domain chairdomain chair,,

www.electroporation.netwww.electroporation.net is is “the best web page” and “the best web page” and he has he has offered offered thethe Newsletter Newsletter reporting on STSMs as an example toreporting on STSMs as an example to other chairs withinother chairs within

thethe BMBS domain.BMBS domain.(Information is from (Information is from NewsletterNewsletter))

What are our activities in COST What are our activities in COST actionaction

The Project has 5 WGsThe Project has 5 WGsWG 1, Basic Mechanisms of Electroporation WG 1, Basic Mechanisms of Electroporation andand Modeling;Modeling; WG 2, Food Processing and WG 2, Food Processing and Pharmaceuticals;Pharmaceuticals;WG 3, Medical ApplicationsWG 3, Medical Applications;;WG 4, Sustainable EnvironmentalWG 4, Sustainable Environmental, , Applications and Biomass Processing;Applications and Biomass Processing;WG 5,WG 5, Technology Development and Technology Development and TransferTransfer

MC membersMC membersAcad. Alexander Petrov, Solid Acad. Alexander Petrov, Solid

State Physics, BASState Physics, BAS

Prof. Bojidar Galutsov, Prof. Bojidar Galutsov, Biological Faculty, SofiaBiological Faculty, Sofia

Договор на тема Договор на тема „Третиране на ракови клетки и „Третиране на ракови клетки и солидни тумори чрез подходящ избор на електрични солидни тумори чрез подходящ избор на електрични импулси” Д0 02/178, импулси” Д0 02/178, финансиран от Националния Фонд финансиран от Националния Фонд

за научни изследвания с кординатор проф., д.б.н. Яна за научни изследвания с кординатор проф., д.б.н. Яна Цонева, И-т по биофизика биомедицинско инженерствоЦонева, И-т по биофизика биомедицинско инженерство, ,

БАНБАН приключи на 30 юни 2012 г. приключи на 30 юни 2012 г.

Title “Treatment of cancer cells and solid tumors by Title “Treatment of cancer cells and solid tumors by suitable choice of electrical pulses”suitable choice of electrical pulses”, , DО 02/178 DО 02/178

The project was scientifically and The project was scientifically and biothecnologically related to a development biothecnologically related to a development of new approaches for electrotreatment of of new approaches for electrotreatment of

tumor cells and solid tumors by tumor cells and solid tumors by

multidisciplinary researchesmultidisciplinary researches

1. Institute of Biophysics and Biomedical 1. Institute of Biophysics and Biomedical Engineering, BASEngineering, BAS

2. Dermatologic Clinic at Specialized Hospital for 2. Dermatologic Clinic at Specialized Hospital for Active Treatment of Oncology, SofiaActive Treatment of Oncology, Sofia3.Institute of Electronics, BAS3.Institute of Electronics, BAS

The new type electroporators Chemipulse III and The new type electroporators Chemipulse III and Chemipulse IV were designed and constructed. In the last Chemipulse IV were designed and constructed. In the last version three elements (electroporator, QRS synhronisator version three elements (electroporator, QRS synhronisator and device for patient current) were incorporated in one and device for patient current) were incorporated in one body. The new instrument is with battery supply, enhanced body. The new instrument is with battery supply, enhanced protection against electrical hazards both for patient and protection against electrical hazards both for patient and the physician. By using the electroporator with QRS the physician. By using the electroporator with QRS synchronization the patients with pacemakers and heard synchronization the patients with pacemakers and heard problems can be treated also. Data collection of the pulse problems can be treated also. Data collection of the pulse parameters and the impedances of treated patients with parameters and the impedances of treated patients with surface tumors could be used to support the optimization surface tumors could be used to support the optimization of the voltage shock setting and the evaluation of the of the voltage shock setting and the evaluation of the procedure effectiveness. The constructed electroporators procedure effectiveness. The constructed electroporators are small and portableare small and portable

Electrochemotherapy of patients included in the studies Electrochemotherapy of patients included in the studies are with clinically and cytologically verified diagnosis are with clinically and cytologically verified diagnosis Carcinoma basocellulare, Carcinoma spinocellulare, Carcinoma basocellulare, Carcinoma spinocellulare, Kaposi sarkoma and Mycosis fungoides, in stage I Kaposi sarkoma and Mycosis fungoides, in stage I (T1N0M0) and the application and study was approved (T1N0M0) and the application and study was approved by the ethics committee of the hospital and all patients by the ethics committee of the hospital and all patients gave written information consent before beginning gave written information consent before beginning treatment. All treatments were carried out at treatment. All treatments were carried out at Dermatologic Clinic at Specialized Hospital for Active Dermatologic Clinic at Specialized Hospital for Active Treatment of Oncology, Sofia. The total number of Treatment of Oncology, Sofia. The total number of successfully treated by electrochemotherapy patients successfully treated by electrochemotherapy patients with device developed during the project was 49, with device developed during the project was 49, number of the treated lesions was 60. The majority of number of the treated lesions was 60. The majority of the lesions are at the visible places at the body and the the lesions are at the visible places at the body and the collection of the material from the lesions for the collection of the material from the lesions for the following of the effect of therapy will lead to the scars following of the effect of therapy will lead to the scars the new method light induced spectroscopy was the new method light induced spectroscopy was

applied.applied.

Dotsinsky, I., Nikolova, B., Peycheva, E., Tsoneva, I.Dotsinsky, I., Nikolova, B., Peycheva, E., Tsoneva, I. 2012,2012, New modality for electrochemotherapy of New modality for electrochemotherapy of surface tumors, surface tumors, Biotechnol. & Biotechnol. Eq. 26, 6, 3402-3406.Biotechnol. & Biotechnol. Eq. 26, 6, 3402-3406.

Николова Николова, , Б.,Б., Цонева, Я., Мудров, Н., Пейчева, Е. 2011, Клинично приложение Цонева, Я., Мудров, Н., Пейчева, Е. 2011, Клинично приложение на електрохимиотерапията: лесен, високо ефективен и безопасен метод за третиране на кожни на електрохимиотерапията: лесен, високо ефективен и безопасен метод за третиране на кожни

тумори. Онкологиятумори. Онкология, 39, 3, 31-35., 39, 3, 31-35.

54 years old woman 1.5 ml bleomycin and 1000 V A)before treatment,B)one week after treatment, C)one month after electrochemotherapy

82 years old woman 2 ml intralesional bleomycin and 1000 V D) before treatment, E) one week later, F) one month follows up

57 years old man 1 ml bleomycin and 900 V G) before treatment, H) 24h after treatment- edema at the site of the treatment, I) 18 months follows up

A) before treatment, B) treatment, C) one week after electrochemotherapy;

A B

C

Advantages of electrochemotherapy

A B

C

A) before treatment, B) one week after treatmentC) one month after

electrochemotherapy;

Electrochemotherapy after surgical operation

Electrochemotherapy

Fundamental studies are related to the increasing Fundamental studies are related to the increasing the amount of the drugs into the cancer cells or the amount of the drugs into the cancer cells or

tumors tumors thus reducing toxicity in normal tissue and thus reducing toxicity in normal tissue and increasing the therapeutic response. Investing the increasing the therapeutic response. Investing the mechanisms of DNA transfer, adhesion behavior mechanisms of DNA transfer, adhesion behavior of cancer cells proliferation and invasiveness we of cancer cells proliferation and invasiveness we

achied higher efficiency in electricity (chemo) achied higher efficiency in electricity (chemo) therapy of tumors.therapy of tumors.

Nikolova, B.,Nikolova, B., Kostadinova, A., Dimitrov, B., Zhelev, Z., Bakalova, R., Aoki, Kostadinova, A., Dimitrov, B., Zhelev, Z., Bakalova, R., Aoki, I., I., Tsoneva, I. 2013, Fluorescent Imaging for Assessment of the Molecular Tsoneva, I. 2013, Fluorescent Imaging for Assessment of the Molecular Mechanisms of Combined Application of Electroporation and Rifampicin on Mechanisms of Combined Application of Electroporation and Rifampicin on HaCaT Cells as a New Therapeutic Approach in Psoriasis, Sensors, 13, 3625-HaCaT Cells as a New Therapeutic Approach in Psoriasis, Sensors, 13, 3625-3634.3634.

Electrotreatment in the presence of antibiotic rifampicin Electrotreatment in the presence of antibiotic rifampicin on HaCat cells (keratinocytes)on HaCat cells (keratinocytes)Fluorescent imaging of Fluorescent imaging of Е-Е-cadxerincadxerin (immunostaining) (immunostaining)A) control, B) HaCat cells, treated with 20 A) control, B) HaCat cells, treated with 20 μμg/mL g/mL rifampicin alone, C) HaCat cells, 24h after rifampicin alone, C) HaCat cells, 24h after electroporation, (D-F) HaCat cells, treated with 20 electroporation, (D-F) HaCat cells, treated with 20 μμg/mL g/mL rifampicin -200V, 500V, 1000V, Magnigication -63xrifampicin -200V, 500V, 1000V, Magnigication -63x

Fluorescent imaging of actin (BODIPYFluorescent imaging of actin (BODIPY558/568558/568phalloidin-phalloidin-labelled)labelled)

1). 4 – PhD students from our Department1). 4 – PhD students from our Department visited the International course visited the International course

Electroporation for MedicineElectroporation for Medicine & & Basic Basic KnowledgeKnowledge, , Applications and TechnologiesApplications and Technologies, , BucharestBucharest, , RumaniaRumania, , OctoberOctober 25-27, 2012 25-27, 2012, ,

financiated by COST action TDfinanciated by COST action TD11041104 2) one PhD student was 3 months in Lubliana, 2) one PhD student was 3 months in Lubliana,

Slovenia, Slovenia, Short-Term Short-Term Scientific Missions (STSMs)Scientific Missions (STSMs), , financiated by financiated by

COST action TDCOST action TD11041104

As a result of our work in the last 4 years the 15 As a result of our work in the last 4 years the 15 papers in journals with impact factor were publeshed papers in journals with impact factor were publeshed (2 are from bilateral work with France and Rumania-(2 are from bilateral work with France and Rumania-participants in COST actionparticipants in COST action TD TD11041104), 4 oral ), 4 oral presentations and 8 posters were presented on the presentations and 8 posters were presented on the international congreses, 5 diploma works and 3 PhD international congreses, 5 diploma works and 3 PhD thesis are defended.thesis are defended.

There is a film about Electrochemotherapy on the There is a film about Electrochemotherapy on the web pages of web pages of Institute of Biophysics and Biomedical Institute of Biophysics and Biomedical Engineering, BASEngineering, BASandand Dermatologic Clinic at Specialized Hospital for Dermatologic Clinic at Specialized Hospital for Active Treatment of Oncology, SofiaActive Treatment of Oncology, Sofia

Българско подзаглавие:Българско подзаглавие:EEлектроиндуцирано въвеждане на конвенционални лектроиндуцирано въвеждане на конвенционални

лекарствени средства и транспортиращи лекарствени средства и транспортиращи наночастици в клетки и туморинаночастици в клетки и тумори

ръководител проекта за съфинансиране - проф. ръководител проекта за съфинансиране - проф. дбн Яна Цонева, член на Мениджмънт комитет от дбн Яна Цонева, член на Мениджмънт комитет от

ИБФБМИ, БАНИБФБМИ, БАН

Electroinduced delivery of conventional drugs and Electroinduced delivery of conventional drugs and transporting nanoparticles in cells and tumorstransporting nanoparticles in cells and tumors( proposal is given on 2.07. 2013, N94-14977)( proposal is given on 2.07. 2013, N94-14977)

1.1.Билатерално сътрудничество със СловенияБилатерално сътрудничество със Словения

2. Билатерално сътрудничество с Румъния2. Билатерално сътрудничество с Румъния

3.3.Билатерално сътрудничество с УкрайнаБилатерално сътрудничество с Украйна

Ин витро изледванияИн витро изледвания- Да се създадат условия за локално увеличение на - Да се създадат условия за локално увеличение на концентрацията на конвенционални противотуморни концентрацията на конвенционални противотуморни лекарства в ракови клетки чрез електропорация. Използване лекарства в ракови клетки чрез електропорация. Използване на наночастици за лечение на тумори.на наночастици за лечение на тумори.- Да се използват възможностите на електропорацията за - Да се използват възможностите на електропорацията за въвеждане на лекарства като милтефозин и еруфозин (нова въвеждане на лекарства като милтефозин и еруфозин (нова група алкилфосфохолини- противотуморни и противо група алкилфосфохолини- противотуморни и противо възпалителни агенти) в керетиноцитни клетки възпалителни агенти) в керетиноцитни клетки HaCaT HaCaT ((ин ин витровитро модел на псориасис). модел на псориасис). - Изучаване на механизмите на действие на електропорацията - Изучаване на механизмите на действие на електропорацията при ко-култури (соматични и неопластични клетки) – Целта е при ко-култури (соматични и неопластични клетки) – Целта е да се изясни как взаимодействат ракови и здрави клетки при да се изясни как взаимодействат ракови и здрави клетки при ръба на тумора след електрохимиотерапия, тъй като макар и ръба на тумора след електрохимиотерапия, тъй като макар и рядко рецидив на заболявнето се появява именно на това рядко рецидив на заболявнето се появява именно на това място.място.- Използване на 2- Използване на 2D D ии 3 3DD култири с оглед доближаване на култири с оглед доближаване на условията на електропорацията на ниво тъкан.условията на електропорацията на ниво тъкан.

Клинични изследванияКлинични изследвания- Да се оптимизират протоколите на - Да се оптимизират протоколите на електрохимиотерапия на кожни тумори и електрохимиотерапия на кожни тумори и оздравителния ефект при тумори, оздравителния ефект при тумори, разположени на различни места по тялото. разположени на различни места по тялото. Да се конкретизира персонализиран подход Да се конкретизира персонализиран подход за лечение на кожни тумори.за лечение на кожни тумори.- Използване на неинвазивни физични - Използване на неинвазивни физични подходи за проследяване на оздравителния подходи за проследяване на оздравителния процес на човешки кожни тумори.процес на човешки кожни тумори.

ЕлектродиЕлектроди1. Нови електроди, съобразени с размера и 1. Нови електроди, съобразени с размера и локализацията на тумора и подсигурени с локализацията на тумора и подсигурени с

подходяща изолоция.подходяща изолоция.