Reusing cell image data for new biological insight (and tool development, and reproducibility)

Examples from my research on how secondary analysis of published cell image data has led to new biological insight.

Transcript

1. Reusing cell image data for new biological insight (and tool development, and reproducibility) @AssafZaritsky https://assafzar.wixsite.com/ascb2017-subgroup
2. Trepat et al. (2009) How (global) coordination emerges from (local) heterogeneous traction forces? Traction Tx (Pa ) Phase Contrast
3. Mechanical information transfer between cells
4. Mechanical information transfer between cells
5. Mechanical information transfer between cells
6. Mechanical information transfer between cells
7. Cell heterogeneity?
8. Suggested model Time Stochastic force exertion transform to directional migration Strain on neighbors coordinate their movement Propagation in time and space to guide groups of cells
9. Measuring traction force, stress and velocity Phase contrast Traction Tx Average normal stress Trepat et al. (2009) Tambe et al. (2011) Serra-Picamal & Conte et al. (2012)
10. Serra-Picamal & Conte et al. (2012) Data from: Serra-Picamal and Conte et al. MDCK cells, N = 4 experiments
11. Serra-Picamal & Conte et al. (2012) Strain rate X-waves (1/4)
12. Alignment of motion and stress
13. Motion-stress alignment Tambe et al. (2011) Trepat & Fredberg. (2011) β −90 ≤∝, ≤ 90 Velocity angle, stress orientation θ 0 ≤ θ ≤ 90 Motion-stress alignment α
14. Plithotaxis Tambe et al. (2011) Trepat and Fredberg (2011) Serra-Picamal and Conte et al. (2012) “tendency for each individual cell within a monolayer to migrate along the local orientation of the maximal principal stress.”
15. Plithotaxis “tendency for each individual cell within a monolayer to migrate along the local orientation of the maximal principal stress.” Tambe et al. (2011) Trepat and Fredberg (2011) Serra-Picamal and Conte et al. (2012)
16. Plithotaxis? “tendency for each individual cell within a monolayer to migrate along the local orientation of the maximal principal stress.” Monolayer edge
17. Plithotaxis? “tendency for each individual cell within a monolayer to migrate along the local orientation of the maximal principal stress.” Monolayer edge Serra-Picamal and Conte et al. (2012)
18. Part I The roles of (global) monolayer geometry versus (local) plithotaxis in inducing motion-stress alignment
19. Motion-Stress Alignment ≠ Plithotaxis Joint distribution % 0 1 Alignment θ
20. No Evidence for Plithotaxis! Monolayer edge
21. ? Observed motion-stress alignment = Global contribution (geometry) + Local contribution (plithotaxis) Components of motion-stress alignment
22. Quantifying the role of monolayer geometry & plithotaxis in motion-stress alignment stress motion Observed alignment Stress Motion Discard pairwise orientation Resampled alignment Random resampling
23. Plithotaxis exists! Geometry > Plithotaxis (N = 4 independent experiments, n = 96 frames in each)
24. The interplay between development of quantitative tools ("hammers“) and identifying open important questions in cell biology ("nails“)
25. What additional information is hidden in data of coupled variables?
26. What do we want to achieve? • Simultaneous investigation of mechanisms that drive global bias and local interactions How? • By modeling the observed agreement between matched variables as the cumulative global and local components Observed colocalization = Global bias + Local interaction
27. Spatio-temporal co-localization of Rac1 and Asef activity in a migrating cell (With Dan Marston, UNC)
28. Part II Properties of cells exhibiting plithotaxis and motion-stress alignment Working hypothesis: enhanced plithotaxis and motion-stress alignment enables more efficient migration during monolayer expansion
29. Physical attributes considered
30. High anisotropy Quantifying plithotaxis for subgroups of cells stress motion Observed alignment Stress Motion Discard pairwise orientation Random resampling Resampled alignment High speed 30
31. Fast cells exhibit elaborated plithotaxis Geometry Plithotaxis plithotaxisall is calculated for all cells. The ratio is calculated for each frame independently (N = 4 independent experiments, n = 96 frames each) = ℎ ℎ , i = Speed, Anisotropy, Strain rate, Stress magnitude Regardless of geometry index
32. Fast cells exhibit elevated directionality
33. Part I & II: Conclusions (local alignment of motion and stress)
34. Part III Intercellular coordination, global alignment of stress and velocity
35. Hypothetical model
36. Hypothetical model
37. Hypothetical model
38. Hypothetical model
39. Hypothetical model
40. From local alignment of motion and stress to intercellular coordination? ?
41. Suggested model Time Stochastic force exertion transform to directional migration Strain on neighbors coordinate their movement Propagation in time and space to guide groups of cells
42. Region-growing segmentation for explicit detection of coordinated migration clusters
43. Associating coordinated stress and motion
44. Coordinated motion is correlated to coordinated stress
45. q p >p*q? Associating coordinated stress and motion
46. Motion- and stress- coordinated clusters are interlinked
47. Cells in coordinated clusters move faster, with enhanced motion-stress alignment Ratioproperty = propertyin propertyout
48. Spatiotemporal analysis
49. Strain-induced motion coordinates cluster’s motility Red: motion clusters, Blue: stress, Magenta: both
50. Strain-induced motion coordinates cluster’s motility Red: motion clusters, Blue: stress Fraction of coordinated cells
51. Das et al. (2015) Data from: Das et al.
52. Stress aligns motion Tight junction proteins play a role in effective transmission of aligned stress to aligned motion
53. Reuse x2
54. Stress aligns motion Tight junction proteins play a role in effective transmission of aligned stress to aligned motion
55. Stress aligns motion Tight junction proteins play a role in effective transmission of aligned stress to aligned motion Could be interesting to look at TJs in WT cells…
56. Part III: Conclusions group plithotaxis Time
57. Coordination migration emerges from cell-cell junctional transmission of mechanical guidance cues Time
58. Part IV Lateral guidance cues (HBEC data is new)
59. Time 1 2 3 Position Protruding cells kymograph
60. Protruding cells kymograph Time 1 2 3 Position
61. Protruding cells kymograph Time 1 2 3 Position
62. Protruding cells kymograph Time 1 2 3 Position
63. Lateral shear-strain waves x-axis
64. Interpretation of diagonal patterns (simplified) Time 1 2 Position
65. Lateral shear-strain waves
66. Rear cells guided to location of shear- strain events at the front
67. Part IV: Conclusions Long range guidance of by leader cells Propagation
68. Summary Seeds of locally aligned motion and stress coordinate collective cell migration Propagation
69. Seeds of Locally Aligned Motion and Stress Coordinate Collective Cell Migration Stochastic force exertion transform to directional migration Strain on neighbors coordinate their movement Propagation in time and space to guide groups of cells
70. Yun-Yu Tseng Angeles Rabadan Xavier Serra- Picamal Xavier Trepat Thanks for sharing your data! Tamal Das Joachim Spatz