Examination arrangement

Course content

Molecular methods are important for diagnosing human diseases and for the follow up of patients during treatment. In addition, molecular methods are important in medical and biological research. In this course the student will be introduced to the use of molecular methods in detection and characterization of driver mutations in human cancer, and to the consequences of such mutations at the cellular level. Methods include cell culture and methods for the measurement of cell growth/-survival., extraction and purification of nucleic acids (DNA and RNA) from human cell lines, gel electrophoresis, polymerase chain reaction (PCR and RT-qPCR), DNA sequencing (Sanger dideoxy method; guided), preparation of protein extracts, detection of epidermal growth factor receptor (EGFR) by Western blot analysis, fixation of cells and in situ immunohistochemical detection of EGFR. Prior to the practical part of the course the student will be presented to guidelines for safe laboratory practices, background theory on human cancer and the role of EGFR, principles for primer and probe design, as well as the use of bioinformatics tools in mutation detection and characterization. Methods for mutation detection and genotyping are important in molecular characterization of cancer, for selecting the most effective first-line treatment, and for detecting molecular alterations in the tumor that may occur during treatment. A range of genotyping methods developed for this purpose will be presented and discussed. Activation of the EGFR pathway (increased EGFR expression, activating EGFR mutation, EGFR gene copy gain) has been implicated in tumorigenesis in non-small cell lung cancer (NSCLC), the most common type of lung cancer. As a result, EGFR has become a key focus for the development of personalized therapy. To identify patients who are most likely to benefit from treatment with EGFR tyrosine kinase inhibitors (TKIs), EGFR mutation testing is being increasingly utilized in clinical practice. Quite commonly, patients who are treated with TKIs will after a while become resistant to the drug. Over 50% of acquired resistance to EGFR TKIs is caused by a mutation in the ATP binding pocket of the EGFR kinase domain involving substitution of a small polar threonine (T) residue with a large nonpolar methionine (M) residue at amino acid position 790 (T790M). The effect of this mutation on cell growth in the presence of TKIs (erlotinib, gefitinib) will be demonstrated.

Learning outcome

Knowledge

The candidate...

• has thorough knowledge in safe laboratory practice
• has thorough knowledge of the principle for Sanger dideoxy DNA sequencing and knowledge of other DNA sequencing methods ("Next generation sequencing")
• has basic knowledge of bioinformatics tools central to molecular medicine
• has good knowledge of PCR methods (PCR, qPCR and RT-qPCR), including basic principles for primer design
• has thorough knowledge of the principle for the Western blot method
• has good theoretical knowledge of in situ immunohistochemical (IHC) detection
• has good theoretical knowledge in animal cell culture

Skills

The candidate...

• can plan and carry out work in the laboratory in such a manner that risks to health and the environment are avoided
• has basic skills in performing Sanger sequencing and sequence analysis using relevant bioinformatic tools
• has basic skills in performing PCR, qPCR and RT-qPCR, including interpretation of PCR results
• has basic skills in performing Western blot analysis
• has basic skills in performing in situ IHC detection
• has good practical skills in cell lab sterile technique and basic practical skills in animal cell culture and measurement of cell viability
• masters data analysis and is able to interpret and critically evaluate own data

General competence

The candidate...

• can analyze relevant academic, professional and research ethical problems
• can apply his/her knowledge and skills in new areas of molecular medicine
• can work with co-students as a team to validate and interpret experimental results from various molecular methods in a biological context, and present and discuss the data obtained as a written scientific paper manuscript

Learning methods and activities

Compulsory lectures, demonstrations and laboratory exercises. Compulsory written assignment (laboratory report). The language of teaching and writing is English. The course has restricted admission and is reserved for students admitted to the Master of Science (MSc) in Molecular Medicine at NTNU.

Compulsory assignments

• Demonstrations
• Lectures

Further on evaluation

Re-take of the exam: If all compulsory assignments are passed, it is sufficient to deliver a new report the following semester. The department may extend the validity of passed compulsory assignments.

Specific conditions

Recommended previous knowledge

MOL3100 (Introduction to Molecular Medicine with Project)

Required previous knowledge

Admission to the Master of Science (MSc) in Molecular Medicine (MSMOLMED) at NTNU.

Course materials

The course material will be announced at the start of the semester.