Periodic Reporting for period 2 - AUTOIGG (AUTOMATED FUNCTIONAL SCREENING OF IgGs FOR DIAGNOSTICS OFNEURODEGENERATIVE DISEASES)
Reporting period: 2020-01-01 to 2023-06-30
The project proposes to organize the exchange of staff of three Academic institutions from Serbia (coordinator), Turkey and Finland, two SMEs from France and Turkey and three TC institutions (two from USA and one from Costa Rica) towards the production of an innovative automated multifunctional device for fluorescence-based diagnostics of neurodegenerative diseases.
The objectives addressed will be:
• Development of experimental cellular models and procedures with immunoglobulins (IgGs) from patient sera as diagnostic and prognostic technologies related to neurodegenerative diseases (particularly based on studies of amyotrophic lateral sclerosis - ALS).
• Defining mark-up characteristics of the standardized in vitro approach for personalized diagnostic protocols.
• Design of a small-scale platform based on automated fluorescence microscopy.
These objectives are based on previous studies on ALS of the coordinators group, however the project also proposes to study the applications on other neuroinflammations and neurodegenerative conditions. This addresses a relevant R & I as well as a socioeconomic medical issue. This research will be chalanged towards application by means of networking that will deal with interesectorial and international exchange of expertise. In addition, three workshops and two training schools will be organized followed by an elaborate dissemination programme. Career plans will be designed for the seconded staff in order to maintain the sustainability of the Action.
The objectives addressed will be:
• Development of experimental cellular models and procedures with immunoglobulins (IgGs) from patient sera as diagnostic and prognostic technologies related to neurodegenerative diseases (particularly based on studies of amyotrophic lateral sclerosis - ALS).
• Defining mark-up characteristics of the standardized in vitro approach for personalized diagnostic protocols.
• Design of a small-scale platform based on automated fluorescence microscopy.
These objectives are based on previous studies on ALS of the coordinators group, however the project also proposes to study the applications on other neuroinflammations and neurodegenerative conditions. This addresses a relevant R & I as well as a socioeconomic medical issue. This research will be chalanged towards application by means of networking that will deal with interesectorial and international exchange of expertise. In addition, three workshops and two training schools will be organized followed by an elaborate dissemination programme. Career plans will be designed for the seconded staff in order to maintain the sustainability of the Action.
WP1. Standardization and benchmarking – overview
Rodent astrocytes as the main cell type were studied for Ca2+ transients (dynamic fluorescent microscopy) induced by ALS IgGs. Same effects were studied in neurons in culture with voltage sensitive dyes
Main results:
- A protocol for automated data acquisition has been designed.
- A software pilot to analyze multiple recordings of Ca2+ transients from neurons in culture is designed and results obtained
- Image segmentation analysis for automated data acquisitions from cells in culture has been accomplished.
- By using fractal analysis and AI approach subcellular characterization of inflammatory markers on astrocytes in culture was accomplished by SEM, supplemented by AFM.
- Autoantibody complementary analysis and testing in clinical environment was performed (autoimmune diseases vs. ALS).
Main training and transfer of knowledge activities thus related to Ca2+ imaging and signal analysis, voltage imaging and machine learning for automated fluorescent signal analysis.
WP2. Pilot platform design – overview
A design and fabrication of a microfluidic chip was performed.
Main results:
- Pilot microfluidic chips were positively tested for flow and solution exchange as well as regarding cell (astroglia) growth and fluorescence-probes' response to stimuli.
- Multiple recordings of Ca2+ transients from neurons in culture were accomplished analyzed by original software and results obtained.
- A preliminary study of synaptic activity was accomplished with voltage sensitive dyes.
- Testing procedures for automated data acquisitions using data samples from Belgrade.
- The optics-on-chip design accomplished includes mobile phone - type micro-camera light aquisition system.
- The lab-on-a-chip brand NIMOCHIP was nationally protected. A patent application is pending.
Main training and transfer of knowledge was in microfluidic biochip fabrication and functional characterization. Software design for data acquisition and analysis.
WP3. Dissemination, communication and networking – overview
• A conference with representatives of 3 RISE projects was organized at ELVESYS.
• Two Training School were organized at UEF as planned
• Three Workshop were organized at YEDITEPE as planned
• Several press releases in the project (1 in Slovenia, 3 in Serbia)
• There were 3 TV appearances of the coordinator.
• A project website was setup at FBUB and a page for the project was designed at the ELVESYS site.
• The project results were presented at about 30 different scientific meetings.
• A showcase session for the project was organized at the Turkish national Neuroscience meeting.
• Two poster presentation on the project results were awarded.
• At a regional Biophysics conference FBUB personnel took part at a pitch event with a talk presenting the project
• In Sept 2018 and 2019 FBUB personnel with seconded staff from ARGENIT and LANOTEC, resp. took part at Researchers Night in Belgrade presenting the project to the general public.
• Two videos were made and posted as YouTube on the project site and FBUB site.
• 8 open access publications with project results were published. Two received press releases in institutional newsletters (both in USA)
• FBUB staff took part in the finals of several accelerator and competition programs (in RS/UK, EU, BE, LV, CN).
• Two national and one EU project were awarded based on knowledge gained in AUTOIGG.
• The innovation NIMOCHIP was preseted in 2023 a the International Technology and Innovation Fair in Belgrade.
WP4. Project management and sustainability – overview
- Kick-off, Mid Term, and 2 Annual meetings were organized in person, and 3 online Annual meetings.
- Management bodies formed were Supervisory board, Secondments Committee, and R & I Council-
- Ethics advisor nominated.
- 2 Amendments have been introduced: 1) new partner at Northwestern University (NWU) substituting partly hosting units from UCHIC, and 2) suspension (>9 months) and overall extension (18 months) of action due to the pandemic.
Rodent astrocytes as the main cell type were studied for Ca2+ transients (dynamic fluorescent microscopy) induced by ALS IgGs. Same effects were studied in neurons in culture with voltage sensitive dyes
Main results:
- A protocol for automated data acquisition has been designed.
- A software pilot to analyze multiple recordings of Ca2+ transients from neurons in culture is designed and results obtained
- Image segmentation analysis for automated data acquisitions from cells in culture has been accomplished.
- By using fractal analysis and AI approach subcellular characterization of inflammatory markers on astrocytes in culture was accomplished by SEM, supplemented by AFM.
- Autoantibody complementary analysis and testing in clinical environment was performed (autoimmune diseases vs. ALS).
Main training and transfer of knowledge activities thus related to Ca2+ imaging and signal analysis, voltage imaging and machine learning for automated fluorescent signal analysis.
WP2. Pilot platform design – overview
A design and fabrication of a microfluidic chip was performed.
Main results:
- Pilot microfluidic chips were positively tested for flow and solution exchange as well as regarding cell (astroglia) growth and fluorescence-probes' response to stimuli.
- Multiple recordings of Ca2+ transients from neurons in culture were accomplished analyzed by original software and results obtained.
- A preliminary study of synaptic activity was accomplished with voltage sensitive dyes.
- Testing procedures for automated data acquisitions using data samples from Belgrade.
- The optics-on-chip design accomplished includes mobile phone - type micro-camera light aquisition system.
- The lab-on-a-chip brand NIMOCHIP was nationally protected. A patent application is pending.
Main training and transfer of knowledge was in microfluidic biochip fabrication and functional characterization. Software design for data acquisition and analysis.
WP3. Dissemination, communication and networking – overview
• A conference with representatives of 3 RISE projects was organized at ELVESYS.
• Two Training School were organized at UEF as planned
• Three Workshop were organized at YEDITEPE as planned
• Several press releases in the project (1 in Slovenia, 3 in Serbia)
• There were 3 TV appearances of the coordinator.
• A project website was setup at FBUB and a page for the project was designed at the ELVESYS site.
• The project results were presented at about 30 different scientific meetings.
• A showcase session for the project was organized at the Turkish national Neuroscience meeting.
• Two poster presentation on the project results were awarded.
• At a regional Biophysics conference FBUB personnel took part at a pitch event with a talk presenting the project
• In Sept 2018 and 2019 FBUB personnel with seconded staff from ARGENIT and LANOTEC, resp. took part at Researchers Night in Belgrade presenting the project to the general public.
• Two videos were made and posted as YouTube on the project site and FBUB site.
• 8 open access publications with project results were published. Two received press releases in institutional newsletters (both in USA)
• FBUB staff took part in the finals of several accelerator and competition programs (in RS/UK, EU, BE, LV, CN).
• Two national and one EU project were awarded based on knowledge gained in AUTOIGG.
• The innovation NIMOCHIP was preseted in 2023 a the International Technology and Innovation Fair in Belgrade.
WP4. Project management and sustainability – overview
- Kick-off, Mid Term, and 2 Annual meetings were organized in person, and 3 online Annual meetings.
- Management bodies formed were Supervisory board, Secondments Committee, and R & I Council-
- Ethics advisor nominated.
- 2 Amendments have been introduced: 1) new partner at Northwestern University (NWU) substituting partly hosting units from UCHIC, and 2) suspension (>9 months) and overall extension (18 months) of action due to the pandemic.
Physiological responses to ALS IgGs in astrocytes and neurons were defined and characterized as neuroinflammatory markers, thus openning the field for novel diagnostic procedures. In addition, networking signaling parameters in ALS IgGs of plated neurons were also analyzed and characterized as a potential innovative diagnostic marker.
Machine learning pilot protocols were designed in order to understand the diagnostic signaling patterns. One PhD was based on these studies. Another master thesis was defended based on the imaging of surface cellular markers and their fractal analysis.
An automated unitary microfluidic device with flow control and measurement protocols is designed as a pilot device. It is meant to be the basic device for the final multicomponent micro-chip system with integral micro-optics.
The research of marketing opportunities has been effectuated and a patent application is pending.
The brand NIMOCHIP is protected nationally. International protection is pending.
SMEs in the project and potential impact
ELVESYS - training in microfluidics design leading to the pilot lab-on-a-chip device.
ARGENIT - automated signal analysis using machine learning was instrumental as the impact on basic studies of cellular dynamic responses to ALS IgGs and automated recording procedure of the pilot device. A publication on the topic and software development prove the efficiency of the project direction towards a commercial and patentable product.
This project also contributed to reach a new marketing niche helping thus academic partners to find entrepreneurs and build a start-up. The company intends to develop advanced diagnostics of ALS and other neuroinflammatory diseases helping patients to obtain early, precise and personalized diagnostics that may also lead ti better therapy design.
Machine learning pilot protocols were designed in order to understand the diagnostic signaling patterns. One PhD was based on these studies. Another master thesis was defended based on the imaging of surface cellular markers and their fractal analysis.
An automated unitary microfluidic device with flow control and measurement protocols is designed as a pilot device. It is meant to be the basic device for the final multicomponent micro-chip system with integral micro-optics.
The research of marketing opportunities has been effectuated and a patent application is pending.
The brand NIMOCHIP is protected nationally. International protection is pending.
SMEs in the project and potential impact
ELVESYS - training in microfluidics design leading to the pilot lab-on-a-chip device.
ARGENIT - automated signal analysis using machine learning was instrumental as the impact on basic studies of cellular dynamic responses to ALS IgGs and automated recording procedure of the pilot device. A publication on the topic and software development prove the efficiency of the project direction towards a commercial and patentable product.
This project also contributed to reach a new marketing niche helping thus academic partners to find entrepreneurs and build a start-up. The company intends to develop advanced diagnostics of ALS and other neuroinflammatory diseases helping patients to obtain early, precise and personalized diagnostics that may also lead ti better therapy design.