Table of Contents
Case 1 – Leber Hereditary Optic Neuropathy (LHON) .......................................................................................... 5
1) What is the detailed function of mitochondria? Especially focus on Complex I-V ................................... 5
Molecular Level and detailed Mechanisms .................................................................................................... 5
Mitochondrial DNA specifics......................................................................................................................... 6
2) What is the hereditary pattern of LHON? And what other types of hereditary patterns apply to general
mitochondrial diseases? Is there anything special about the genetic segregation or ageing development? ........ 7
Maternal Inheritance....................................................................................................................................... 8
Sporadic Diseases ........................................................................................................................................... 8
Mendelian Diseases ........................................................................................................................................ 9
Ageing-Related somatic accumulation ........................................................................................................... 9
3) How does LHON progress and what does it affect within the human body? And what are the long-term
and global consequences? How do other more general mitochondrial diseases progress? ............................... 10
Pathophysiology ........................................................................................................................................... 10
Clonal expansion of mtDNA ........................................................................................................................ 10
Mitophagy .................................................................................................................................................... 11
Mitochondrial Haplotypes ............................................................................................................................ 11
4) How can mitochondrial diseases be diagnosed and/or can they be prognosed? Are there any preventative
measures that can be taken against LHON or are there ways to relieve or slow down the progression of
LHON? ............................................................................................................................................................ 11
Treatments .................................................................................................................................................... 12
Genetic Counselling ..................................................................................................................................... 13
Therapies under investigation....................................................................................................................... 13
Case 2 – Mitochondrial Replacement Theories .................................................................................................... 15
1) What type of Mitochondrial Replacement Technique (MRT) approaches are available and what are their
success rates? In what other cases can it be used? ............................................................................................ 15
Extended Applications for MRT .................................................................................................................. 17
2) What is IVF? ............................................................................................................................................ 18
3) What are the legal implications of MRT in the Netherlands? .................................................................. 18
Ethical and Legal issues in Mitochondrial Transfer – Newson, Wilkinson, and Wrigley ............................ 19
4) What are the ethical concerns of MRT in the Netherlands? ..................................................................... 20
Donor Anonymity......................................................................................................................................... 21
Lecture-1 Mito-1 - Understanding Cellular Energy Production ............................................................................ 22
Electron Transport Mechanism .................................................................................................................... 23
CHALLENGES ............................................................................................................................................ 24
Lecture-2 Mito-2 - Understanding Cellular Energy Production ............................................................................ 25
CHALLENGES MITOCHONDRIAL DISEASE DIAGNOSIS.................................................................. 27
Lecture-3 Mito-3 - Mitochondrial Disorders......................................................................................................... 28
Lecture-4 Mito-4 Q&A ......................................................................................................................................... 30
Case 3 – Post Traumatic Stress Disorder .............................................................................................................. 31
1) What are the causes/risks of PTSD? Focus on both biological ones and life events ................................ 31
1
, Prevalence .................................................................................................................................................... 31
Course .......................................................................................................................................................... 31
Risk Factors .................................................................................................................................................. 31
2) What brains regions, neurological pathways, and neurotransmitters are involved? ................................. 32
Cognitive Behavioural Methods ................................................................................................................... 32
Mechanisms and Pathophysiology ............................................................................................................... 33
3) What symptoms can PTSD patients have? ............................................................................................... 38
4) How is PTSD diagnosed? ........................................................................................................................ 39
5) What are the possible treatment options? Therapy and Medication ......................................................... 41
Psychotherapy .............................................................................................................................................. 41
Pharmacotherapy .......................................................................................................................................... 41
6) Research-related problems (Lecture based) ............................................................................................. 42
Lecture-5 Neuro-1 – PTSD (Implemented in case 3) ............................................................................................ 43
Case 4 – Epilepsy and Schizophrenia ................................................................................................................... 46
1) What are the symptoms of Schizophrenia and Temporal Lobe Epilepsy (TLE)? .................................... 46
Schizophrenia ............................................................................................................................................... 46
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 47
Similarities among both................................................................................................................................ 48
2) What are the triggers and causes (risk factors, genetics, aetiology) of Schizophrenia and Temporal Lobe
Epilepsy (TLE)? ............................................................................................................................................... 49
Schizophrenia ............................................................................................................................................... 49
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 50
Similarities among both................................................................................................................................ 50
3) What are the molecular mechanisms/pathways for Schizophrenia and Temporal Lobe Epilepsy (TLE)?
51
Schizophrenia ............................................................................................................................................... 51
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 52
Similarities among both................................................................................................................................ 55
4) What are diagnostic methods of Schizophrenia and Temporal Lobe Epilepsy (TLE)? ........................... 56
Schizophrenia ............................................................................................................................................... 56
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 56
Similarities among both................................................................................................................................ 57
5) What are treatments and medications for Schizophrenia and Temporal Lobe Epilepsy (TLE)? ............. 57
Schizophrenia ............................................................................................................................................... 57
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 58
Similarities among both................................................................................................................................ 59
Case 5 – Too Fat or Not Too Fat ........................................................................................................................... 60
1) What is defined as metabolically healthy individual? .............................................................................. 60
Stress ............................................................................................................................................................ 61
Obesity ......................................................................................................................................................... 61
Sedentary Lifestyle ....................................................................................................................................... 61
2
, Pathophysiology ........................................................................................................................................... 61
2) How and where is fat stored? Visceral and sub cutaneous Fat, and how do these develop? (Storage
depot; Storing vs Proliferating and creating new adipocytes) .......................................................................... 61
Visceral Fat .................................................................................................................................................. 62
Epicardial Fat ............................................................................................................................................... 63
Subcutaneous Fat.......................................................................................................................................... 63
Marrow Fat ................................................................................................................................................... 63
Ectopic Fat.................................................................................................................................................... 63
Brown Fat ..................................................................................................................................................... 65
Physiology .................................................................................................................................................... 65
3) Why is BMI a bad index for identifying health? (bodybuilder?) (FAT MASS VS FAT FREE MASS) . 65
Alternatives .................................................................................................................................................. 66
Case 6 – Foye Grass (… Ufff … Foie Gras, ya happy? Haha) ............................................................................. 67
1) What is Non-Alcoholic Fatty Liver Disease (NAFLD)? How does it develop? What are the different
stages?............................................................................................................................................................... 67
2) What are the symptoms and diagnostic methods for NAFLD/Fatty Liver? (And what are possible
treatments) ........................................................................................................................................................ 69
3) What are the causes and risk factors for NAFLD? ................................................................................... 70
4) What is the most important organ in Type-II Diabetes and what is inter-organ crosstalk? ..................... 71
Focussing on Liver-Muscle Cross-Talk ....................................................................................................... 73
Lecture-Doesn’t Matter Metabolism-1 - Metabolism ........................................................................................... 74
Metabolically Healthy and Unhealthy Picture .................................................................................................. 78
Causation Workshop ............................................................................................................................................. 80
Assignment-1 .................................................................................................................................................... 80
a) The age of the person and the extent of the physical damage?................................................................. 80
b) The height of the fall and the extent of the physical damage? ................................................................. 81
c) The age of the person and the height of the fall? ...................................................................................... 82
a) The age of the person and the extent of the physical damage?................................................................. 82
b) The extent of the physical damage and the height of the fall? ................................................................. 82
Assignment-2 .................................................................................................................................................... 82
Question A: Design an in vitro experiment to investigate whether Selenoprotein P improves glucose uptake
in muscle cells. Does this experiment prove causality? Why (not?) ............................................................ 82
Question B: Design an experiment to investigate whether there is a dose response relationship between the
concentration of Selenoprotein P and glucose uptake. Does this experiment prove causality? Why (not)? 83
Question C: A researcher has 2 groups of mice. One group of normal mice, and one group of mice that is
injected with Selenoprotein P. The investigator then measures glucose uptake. Does this experiment give
any information related to causality? Explain why (not). ............................................................................. 83
Question D: A researcher has 2 groups of mice. One group of normal mice, and one group of mice that is
genetically modified. In this latter group, the Selenoprotein P receptor is knocked-out. The investigator
places all mice on a high fat diet and investigates whether they develop insulin resistance. Does this
experiment give any information related to causality? Explain why (not). .................................................. 83
Question E: A researcher measures glucose uptake in a large group of mice. He then places the mice on an
exercise regime. This results in decreased levels of selenoprotein P, and glucose uptake improves. What
does this experiment tell us about causality? ................................................................................................ 83
3
, Assignment-2 .................................................................................................................................................... 83
Question A: Injection of recombinant fetuin-A in mice decreases insulin sensitivity. ................................. 83
Question B: Fetuin B is elevated in type 2 diabetes patients compared with lean, normoglycemic
individuals. ................................................................................................................................................... 84
Question C: ANGPTL4 overexpression in mice reduces adipose tissue weight and leads to liver steatosis
and elevated plasma levels of triglycerides. ................................................................................................. 84
Question D: Serum FGF21 levels reflect the degree of steatosis in patients with diabetes .......................... 84
Presentation Notes – Polar Body Genome Transfer .............................................................................................. 84
1) Introduction .............................................................................................................................................. 84
2) Hypothesis ............................................................................................................................................... 85
3) Materials and Methods ............................................................................................................................. 85
Pyrosequencing ............................................................................................................................................ 86
Pitch Presentation Notes – Prime Editing ............................................................................................................. 86
Mentor Meeting Card ............................................................................................................................................ 87
Workshop PTSD ................................................................................................................................................... 88
Cell Cultures ..................................................................................................................................................... 88
Drug Treatment................................................................................................................................................. 88
Whatsapp Answers ................................................................................................................................................ 90
4
Case 1 – Leber Hereditary Optic Neuropathy (LHON) .......................................................................................... 5
1) What is the detailed function of mitochondria? Especially focus on Complex I-V ................................... 5
Molecular Level and detailed Mechanisms .................................................................................................... 5
Mitochondrial DNA specifics......................................................................................................................... 6
2) What is the hereditary pattern of LHON? And what other types of hereditary patterns apply to general
mitochondrial diseases? Is there anything special about the genetic segregation or ageing development? ........ 7
Maternal Inheritance....................................................................................................................................... 8
Sporadic Diseases ........................................................................................................................................... 8
Mendelian Diseases ........................................................................................................................................ 9
Ageing-Related somatic accumulation ........................................................................................................... 9
3) How does LHON progress and what does it affect within the human body? And what are the long-term
and global consequences? How do other more general mitochondrial diseases progress? ............................... 10
Pathophysiology ........................................................................................................................................... 10
Clonal expansion of mtDNA ........................................................................................................................ 10
Mitophagy .................................................................................................................................................... 11
Mitochondrial Haplotypes ............................................................................................................................ 11
4) How can mitochondrial diseases be diagnosed and/or can they be prognosed? Are there any preventative
measures that can be taken against LHON or are there ways to relieve or slow down the progression of
LHON? ............................................................................................................................................................ 11
Treatments .................................................................................................................................................... 12
Genetic Counselling ..................................................................................................................................... 13
Therapies under investigation....................................................................................................................... 13
Case 2 – Mitochondrial Replacement Theories .................................................................................................... 15
1) What type of Mitochondrial Replacement Technique (MRT) approaches are available and what are their
success rates? In what other cases can it be used? ............................................................................................ 15
Extended Applications for MRT .................................................................................................................. 17
2) What is IVF? ............................................................................................................................................ 18
3) What are the legal implications of MRT in the Netherlands? .................................................................. 18
Ethical and Legal issues in Mitochondrial Transfer – Newson, Wilkinson, and Wrigley ............................ 19
4) What are the ethical concerns of MRT in the Netherlands? ..................................................................... 20
Donor Anonymity......................................................................................................................................... 21
Lecture-1 Mito-1 - Understanding Cellular Energy Production ............................................................................ 22
Electron Transport Mechanism .................................................................................................................... 23
CHALLENGES ............................................................................................................................................ 24
Lecture-2 Mito-2 - Understanding Cellular Energy Production ............................................................................ 25
CHALLENGES MITOCHONDRIAL DISEASE DIAGNOSIS.................................................................. 27
Lecture-3 Mito-3 - Mitochondrial Disorders......................................................................................................... 28
Lecture-4 Mito-4 Q&A ......................................................................................................................................... 30
Case 3 – Post Traumatic Stress Disorder .............................................................................................................. 31
1) What are the causes/risks of PTSD? Focus on both biological ones and life events ................................ 31
1
, Prevalence .................................................................................................................................................... 31
Course .......................................................................................................................................................... 31
Risk Factors .................................................................................................................................................. 31
2) What brains regions, neurological pathways, and neurotransmitters are involved? ................................. 32
Cognitive Behavioural Methods ................................................................................................................... 32
Mechanisms and Pathophysiology ............................................................................................................... 33
3) What symptoms can PTSD patients have? ............................................................................................... 38
4) How is PTSD diagnosed? ........................................................................................................................ 39
5) What are the possible treatment options? Therapy and Medication ......................................................... 41
Psychotherapy .............................................................................................................................................. 41
Pharmacotherapy .......................................................................................................................................... 41
6) Research-related problems (Lecture based) ............................................................................................. 42
Lecture-5 Neuro-1 – PTSD (Implemented in case 3) ............................................................................................ 43
Case 4 – Epilepsy and Schizophrenia ................................................................................................................... 46
1) What are the symptoms of Schizophrenia and Temporal Lobe Epilepsy (TLE)? .................................... 46
Schizophrenia ............................................................................................................................................... 46
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 47
Similarities among both................................................................................................................................ 48
2) What are the triggers and causes (risk factors, genetics, aetiology) of Schizophrenia and Temporal Lobe
Epilepsy (TLE)? ............................................................................................................................................... 49
Schizophrenia ............................................................................................................................................... 49
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 50
Similarities among both................................................................................................................................ 50
3) What are the molecular mechanisms/pathways for Schizophrenia and Temporal Lobe Epilepsy (TLE)?
51
Schizophrenia ............................................................................................................................................... 51
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 52
Similarities among both................................................................................................................................ 55
4) What are diagnostic methods of Schizophrenia and Temporal Lobe Epilepsy (TLE)? ........................... 56
Schizophrenia ............................................................................................................................................... 56
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 56
Similarities among both................................................................................................................................ 57
5) What are treatments and medications for Schizophrenia and Temporal Lobe Epilepsy (TLE)? ............. 57
Schizophrenia ............................................................................................................................................... 57
Temporal Lobe Epilepsy (TLE) ................................................................................................................... 58
Similarities among both................................................................................................................................ 59
Case 5 – Too Fat or Not Too Fat ........................................................................................................................... 60
1) What is defined as metabolically healthy individual? .............................................................................. 60
Stress ............................................................................................................................................................ 61
Obesity ......................................................................................................................................................... 61
Sedentary Lifestyle ....................................................................................................................................... 61
2
, Pathophysiology ........................................................................................................................................... 61
2) How and where is fat stored? Visceral and sub cutaneous Fat, and how do these develop? (Storage
depot; Storing vs Proliferating and creating new adipocytes) .......................................................................... 61
Visceral Fat .................................................................................................................................................. 62
Epicardial Fat ............................................................................................................................................... 63
Subcutaneous Fat.......................................................................................................................................... 63
Marrow Fat ................................................................................................................................................... 63
Ectopic Fat.................................................................................................................................................... 63
Brown Fat ..................................................................................................................................................... 65
Physiology .................................................................................................................................................... 65
3) Why is BMI a bad index for identifying health? (bodybuilder?) (FAT MASS VS FAT FREE MASS) . 65
Alternatives .................................................................................................................................................. 66
Case 6 – Foye Grass (… Ufff … Foie Gras, ya happy? Haha) ............................................................................. 67
1) What is Non-Alcoholic Fatty Liver Disease (NAFLD)? How does it develop? What are the different
stages?............................................................................................................................................................... 67
2) What are the symptoms and diagnostic methods for NAFLD/Fatty Liver? (And what are possible
treatments) ........................................................................................................................................................ 69
3) What are the causes and risk factors for NAFLD? ................................................................................... 70
4) What is the most important organ in Type-II Diabetes and what is inter-organ crosstalk? ..................... 71
Focussing on Liver-Muscle Cross-Talk ....................................................................................................... 73
Lecture-Doesn’t Matter Metabolism-1 - Metabolism ........................................................................................... 74
Metabolically Healthy and Unhealthy Picture .................................................................................................. 78
Causation Workshop ............................................................................................................................................. 80
Assignment-1 .................................................................................................................................................... 80
a) The age of the person and the extent of the physical damage?................................................................. 80
b) The height of the fall and the extent of the physical damage? ................................................................. 81
c) The age of the person and the height of the fall? ...................................................................................... 82
a) The age of the person and the extent of the physical damage?................................................................. 82
b) The extent of the physical damage and the height of the fall? ................................................................. 82
Assignment-2 .................................................................................................................................................... 82
Question A: Design an in vitro experiment to investigate whether Selenoprotein P improves glucose uptake
in muscle cells. Does this experiment prove causality? Why (not?) ............................................................ 82
Question B: Design an experiment to investigate whether there is a dose response relationship between the
concentration of Selenoprotein P and glucose uptake. Does this experiment prove causality? Why (not)? 83
Question C: A researcher has 2 groups of mice. One group of normal mice, and one group of mice that is
injected with Selenoprotein P. The investigator then measures glucose uptake. Does this experiment give
any information related to causality? Explain why (not). ............................................................................. 83
Question D: A researcher has 2 groups of mice. One group of normal mice, and one group of mice that is
genetically modified. In this latter group, the Selenoprotein P receptor is knocked-out. The investigator
places all mice on a high fat diet and investigates whether they develop insulin resistance. Does this
experiment give any information related to causality? Explain why (not). .................................................. 83
Question E: A researcher measures glucose uptake in a large group of mice. He then places the mice on an
exercise regime. This results in decreased levels of selenoprotein P, and glucose uptake improves. What
does this experiment tell us about causality? ................................................................................................ 83
3
, Assignment-2 .................................................................................................................................................... 83
Question A: Injection of recombinant fetuin-A in mice decreases insulin sensitivity. ................................. 83
Question B: Fetuin B is elevated in type 2 diabetes patients compared with lean, normoglycemic
individuals. ................................................................................................................................................... 84
Question C: ANGPTL4 overexpression in mice reduces adipose tissue weight and leads to liver steatosis
and elevated plasma levels of triglycerides. ................................................................................................. 84
Question D: Serum FGF21 levels reflect the degree of steatosis in patients with diabetes .......................... 84
Presentation Notes – Polar Body Genome Transfer .............................................................................................. 84
1) Introduction .............................................................................................................................................. 84
2) Hypothesis ............................................................................................................................................... 85
3) Materials and Methods ............................................................................................................................. 85
Pyrosequencing ............................................................................................................................................ 86
Pitch Presentation Notes – Prime Editing ............................................................................................................. 86
Mentor Meeting Card ............................................................................................................................................ 87
Workshop PTSD ................................................................................................................................................... 88
Cell Cultures ..................................................................................................................................................... 88
Drug Treatment................................................................................................................................................. 88
Whatsapp Answers ................................................................................................................................................ 90
4
