12946 - Molecular Biology

Learning outcomes

At the end of the Course, Students will know:

1. The biophysical dynamics of cellular microtubules, within the context of the most advanced views in intra-, inter-cellular communication, and biomolecular recognition.
2. The nuclear trafficking of signaling molecules and transcription factors.
3. The main activation pathways of transcription factors and their nuclear mechanisms of action at the molecular and nanomechanical level, with particular reference to the processes of: gene expression, cell proliferation and differentiation.
4. The role of epigenetics in the modulation of cellular homeostasis, highlighting the molecular mechanisms responsible for stem cell pluripotency. In this regard, particular emphasis will be placed to the use of physical energies (including mechanical vibration and electromagnetic radiation) to afford efficient in situ reprogramming of tissue-resident stem cells, paving the way to a Regenerative Medicine without the needs for (stem)cell or tissue transplantation.

Course contents

The Course is divided into a total of 4 lessons. The topics covered for each lesson are shown below. The description of these topics also intends to provide an indication of the contribution of each lesson to the knowledge and skills to be achieved, as it is underlined in the Biomedical Implications.

1. Analysis of the physical dynamics of cellular microtubules. Microtubules are entities capable of mechanical and electromagnetic oscillations. Methods for studying the oscillations of microtubules at the nanomechanical and electromagnetic level: atomic force microscopy (AFM) and hyperspectral imaging (HSI). Microtubules as a network of oscillators capable of synchronization and coordinated migration (swarming). The dynamics of microtubules in biomolecular recognition processes. The "memory switching" properties inherent in individual microtubules. Biomedical implications: microtubuli behave as a bioelectronic circuit capable of generating essential information in cell memory and connectedness.
2. Nuclear trafficking of molecular signals. The nuclear pore complex. RAN-TC and Karyopherins. The nuclear import of transcription factors and protein kinases: prelude to epigenetic regulation, chromatin and gene transcription remodeling.

   Transcriptional regulation. Physical remodeling of chromatin: DNA bending and DNA loops to guide the movement and sliding speed of the RNA polymerase complex. Transcription factors: Zinc Fingers, Nuclear Hormone Receptors (NHR), Homeodomains, and Leucine Zippers. Implications in the differentiation and maintenance of cell identity. Transcription factors act as actuators of mechanical forces inside the transcriptional machinery. Biomedical implications: The nucleus as a "portal" for the integration of essential biophysical signals in the regulation of cell biology.The nanomechanics of transcriptional processes is one of the new frontiers of epigenetic regulation and regenerative medicine.

3. The stromal-vascular niche: a nanotopography where chemical and physical signals come together to modulate the biology of stem cells. Use of weak mechanical forces for the processing of autologous adipose tissue into micro-fragmented preparations capable of preserving the stem component within the context of an intact stromal-vascular niche. Biomedical implications: development of adipose tissue derivatives ready for autologous use in regenerative medicine. Our experience in orthopedic, vascular, and dental regenerative medicine.
4. Use of physical energies to afford efficient in situreporgramming of human adult stem and somatic cells. The development of a Regenerative Medicine without (stem)cell and tissue transplantation.
   • Ad hoc conveyed radioelectric fields for the modulation of stem cell fate. Our experience in Regenerative Medicine: from the reversibility of the stem cell senescence process, to the induction of cardiogenesis, neurogenesis, vasculogenesis and skeletal myogenesis. Use of electromagnetic fields for the direct reprogramming of human adult somatic cells.
   • Subsonic and acoustic mechanical vibrations for the modulation of the differentiating potential of human adult stem cells. Our experience in deciphering vibrational nanomechanical patterns by Atomic Force Microscopy (AFM) and Hyperspectral Imaging (HSI) in stem cells, and developing mechanical actuators to induce specific stem cell differentiation.

Biomedical Implications: the chance of using the diffusive features of physical energies, including mechanical vibrations and electromagnetic radiation (electromagnetic fields and light), to enhance the self- healing potential afforded by tissue-resident stem cells.

Readings/Bibliography

Readings and bibliography will be provided throughout the Course, during each lesson, stored and made available in the IOL section.

Teaching methods

• Computer assisted presentations.
• Discussion of experimental findings.
• Seminars, particularly focused on novel approaches in Regenerative Medicine.
• Presentation and discussion of main findings and major conclusions from International Meetings coherent with the aim, contents and outcomes of the Course.

Assessment methods

• Computer assisted presentations.
• Discussion at the end of each lesson.
• Discussion of experimental findings.
• Questions and highlights.
• Seminars, particularly focused on novel approaches in Regenerative Medicine.
• The final exam will focus on topics covered during the lessons, and will include a computer presentation of a short paper, arranged by the candidate, in suitable format (PPTX, Keynote, pdf).
• The achievement by the Student of an organic vision of the themes developed during the Course, together with a capacity for critical analysis of the topics learned, and the development of a personal narrative, will be evaluated with marks of excellence.
• The manifestation of a mostly mnemonic/descriptive knowledge of the topics covered in class, together with non-articulated skills of analysis and synthesis, developed through a narrative that is not always precise and correct, will lead to a fair exam grade.
• Evidence of training gaps, in the presence of deficiencies in the analysis and synthesis processes of the topics required during the exam, will lead to a sufficiency judgment, provided that the Student expresses evidence for a minimal knowledge of the discussed subjects.
• Significant training gaps, together with inappropriate language and lack of orientation in the context of the topics covered, as well as of the training and bibliographic materials offered during the Course, could only be evaluated negatively.

Teaching tools

• Power point presentations.
• Critical discussion of personal research and review articles.
• Critical readings of studies published by other Authors.
• Presentation of movies form international meetings.

Office hours

See the website of Carlo Ventura