molecular cell biology (bio 5068) · (3) cystogenesis causes kidney fibrosis and scarring bergmann...
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Ciliopathies and Centrosomopathies
Molecular Cell Biology (Bio 5068)
@MahjoubLab
Moe R. Mahjoub Department of Medicine (Nephrology), Cell Biology & Physiology
Washington University in St Louis
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Human Ciliopathies: Diseases caused by defective ciliary function
Sjogren
Leber's congenital amaurosis
Senior-Loken
Polycystic kidney disorder
Sensenbrenner syndrome
von Hippel-Lindau
Short rib polydactyly disorders
Joubert syndrome
Nephronophthisis
Meckel-Gruber
Alstrom
Bardet-Biedl
Primary cilia dyskinesia
eye
kidney
liver
skeletal
CNS
laterality
obesity
respiratorytract
Image: Susan Dutcher
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Outline:
! Motile Ciliopathies • Left-Right asymmetry • Primary Ciliary Dyskinesia
! Sensory Ciliopathies • Polycystic Kidney Disease • Retinal degeneration
! Centrosomopathies • Microcephaly
• Cancer
Reiter and Leroux, Nature Reviews – MCB (2018)
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Nodal cilia, a rare type of motile cilium…
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Symmetry breaking by nodal cilia-mediated flow
D. Grimes, Development (2019)
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Symmetry breaking by nodal cilia-mediated flow
Yaun et al, Current Biology (2015)
Zebrafish L-R organizer
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Nodal cilia dysfunction causes organ laterality defects
• Rare congenital disease – affects roughly 1 in 10,000 live births
• Inheritance: either autosomal recessive (most commonly), autosomal dominant, or X-linked.
• Mutations can be in ciliary genes (motility or signaling), or in morphogenesis pathways
• Patients with situs inversus totalis can live normally with few symptoms
• Heterotaxy syndrome (situs ambiguus) – associated with heart, lung, and other organ defects that can be fatal
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Diseases of the airway: Primary Ciliary Dyskinesia (PCD)
Ciliated cells
Secretory cells (mucus, surfactant)
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The Mucociliary Escalator
https://kids.frontiersin.org
J. Whitsett, Annals of the American Thoracic Society (2018)
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Primary Ciliary Dyskinesia (PCD)
https://kids.frontiersin.org Zhou-Suckow et al, Cell and Tissue Research (2017)
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Primary Ciliary Dyskinesia (PCD)
https://kids.frontiersin.org Mirra et al, Frontiers in Pediatrics (2017)
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Primary Ciliary Dyskinesia (PCD)
Mirra et al, Frontiers in Pediatrics (2017)
• Rare, inherited, autosomal recessive (mostly), although example of X-linked inheritance exists
• Congenital disease – 1 in 20,000 live births
• At least 40 genes identified so far (but almost half the genes are unknown)
• Mutations mostly disrupt ciliary dynein motor preassembly and trafficking into cilia
• ~50% of patients also display organ laterality defects
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Mutations in PCD and impact on cilia ultrastructure
Mirra et al, Frontiers in Pediatrics (2017) Horani and Ferkol, Chest (2018)
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Treatment regiments in PCD patients
Guan et al, Current Allergy and Asthma Reports (2018)
In addition to this inherited genetic disease, there are also acquired cilia-associated airway diseases (e.g. COPD, asthma)
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Outline:
! Motile Ciliopathies • Left-Right asymmetry • Primary Ciliary Dyskinesia
! Sensory Ciliopathies • Polycystic Kidney Disease • Retinal degeneration
! Centrosomopathies • Microcephaly
• Cancer
Reiter and Leroux, Nature Reviews – MCB (2018)
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Sensory Ciliopathies: Polycystic Kidney Disease
• Most common ciliopathy – 1 in 400 people
• Most common inherited monogenic kidney disease
• Autosomal Dominant (ADPKD) • Mutations in 2 genes - PKD1 (85%) and PKD2 (10%)
• Slow, degenerative disease
• Results in end-stage kidney failure in the 50-60’s
• Autosomal Recessive (ARPKD) • Rare (1 in 20,000)
• Mutations in a single gene – PKHD1
• Rapid onset
• Both ADPKD and ARPKD are associated with extra-renal manifestations (e.g. liver cysts)
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Mechanosensory role of the primary cilium in nephrons
O’Connor et al, Cilia (2013)
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Mechanosensory role of the primary cilium in nephrons
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Molecular mechanisms of renal cystogenesis
Bergmann et al, Nat. Rev. Dis. Primers (2018)
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Cellular mechanisms of renal cystogenesis
Bergmann et al, Nat. Rev. Dis. Primers (2018)
(1)
(2)
(3)
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Cystogenesis causes kidney fibrosis and scarring
Bergmann et al, Nat. Rev. Dis. Primers (2018)
Xue and Mei, Renal Fibrosis: Mechanisms and Therapies (2019)
Defective ciliary
signaling
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Sensory Ciliopathies: Retinal Degeneration
The outer segment of photoreceptor cells is a highly
modified sensory cilium
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Mechanism of ciliary assembly and protein trafficking in photoreceptors
May-Simera et al, Progress in Retinal and Eye Research (2017)
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Mechanism of ciliary assembly and protein trafficking in photoreceptors
Nemet et al, Progress in Molecular Biology and Translational Science (2015)
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Patho-mechanism of retinal degeneration
• Mutations in ~250 genes identified so far
• Most common inherited disease is Retinitis pigmentosa (affects 1 in 3000 people)
• Symptoms begin in the 20’s - patients typically begin to develop night blindness, which turns into tunnel vision and ultimately blindness.
Mutations disrupt: • Centriole formation (genes essential for biogenesis) • Centriole appendages (docking of mother centriole
disrupted) • IFT genes (trafficking into cilia abnormal) • Transition zone (aberrant ciliary protein composition)
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Patho-mechanism of retinal degeneration
May-Simera et al, Progress in Retinal and Eye Research (2017)
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Outline:
! Motile Ciliopathies • Left-Right asymmetry • Primary Ciliary Dyskinesia
! Sensory Ciliopathies • Polycystic Kidney Disease • Retinal degeneration
! Centrosomopathies • Microcephaly
• Cancer
Reiter and Leroux, Nature Reviews – MCB (2018)
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Autosomal Recessive Primary Microcephaly
• Rare - affects 1 in 10,000-250,000 people • The majority (80%) of mutations are in centrosome genes
Mutations impact: • Centriole formation (genes essential for biogenesis) • Pericentriolar material (PCM) – microtubule organization • Mitosis
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Cellular mechanism of pathogenesis
• Centrosomes control fate decisions during asymmetric cell division
• Mutations in centrosome genes disrupt this process
“Older” centrosome “Younger” centrosome
Pelletier and Yamashita (2012)
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Cellular mechanism of pathogenesis
• Neural progenitor cells (NPCs) undergo a series of symmetric proliferative divisions during early neurogenesis to expand the NPC pool.
• These cells then switch to an asymmetric mode of division that
generates neurons and maintains the NPC pool throughout the later stages of neurogenesis (top).
• Defects in centrosome-related microcephaly (CRM) genes can
disrupt neurogenic divisions, resulting in loss of NPCs by: o premature differentiation due to spindle mis-orientation o cell cycle delays (mitotic)
o activation of apoptotic pathways due to failure to satisfy the spindle assembly checkpoint
o Chromosome missegregation and aneuploidy .
• Collectively, these defects result in reduction of the abundance of neurons, causing smaller brain size
O’Neill et al, Molecular Biology of the Cell (2018)
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Abnormal centrosome biogenesis and cancer
Overduplication Cytokinesis failure
Cell fusion Viral infection
Boveri, 1914: extra centrosomes drive genome instability in tumors
Galeotti, 1896
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Abnormal centrosome biogenesis and cancer
Cosensa and Kramer, Chromosome Research, 2015
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Centrosome clustering helps cancer cells survive
P30 P45 P75
Milunovic-Jevtic et al, 2016
Centrosome Clustering
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Centrosome clustering helps cancer cells survive
Cosensa and Kramer, Chromosome Research, 2015
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Centrosome-dependent mechanism of tumor formation
Arnandis et al, Dev Cell, 2018
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Centrosome-dependent mechanism of tumor formation
• The majority of solid tumors (>80%) contain cells with extra centrosomes
• But, these cells only make up a small fraction of the tumor population (between 10-20% of cells).
• How can a small population of cells with abnormal centrosome number drive tumor growth?
• Cause genomic instability in daughter cells after cell division
• Induce proliferation of neighboring cells via paracrine signaling
• Promote invasivness by increasing microtubule number emanating from the centrosome
Arnandis et al, Dev Cell, 2018
Nigg and Holland, Nature Reviews – Molecular Cell Biology (2018)
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P30 P45 P75
Milunovic-Jevtic et al, 2018
Genome instability Ciliary defects Enhanced secretion
Targeting cells with centrosome amplification as a therapeutic strategy
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Inhibition of centrosome clustering in vitro
P30 P45 P75
Mariappan et al, EMBO Reports (2018)
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Inhibition of centrosome clustering in vivo
P30 P45 P75
Mariappan et al, EMBO Reports (2018)