Presenter: Margo Emont, PhD. Instructor, Beth Israel Deaconess Medical Center/Harvard Medical School Abstract White adipose tissue, once regarded as morphologically and functionally bland, is now recognized to be dynamic, plastic and heterogenous, and is involved in a wide array of biological processes including energy homeostasis, glucose and lipid handling, blood pressure control and host defense. High-fat feeding and other metabolic stressors cause marked changes in adipose morphology, physiology and cellular composition, and alterations in adiposity are associated with insulin resistance, dyslipidemia and type 2 diabetes. Here we provide detailed cellular atlases of human and mouse subcutaneous and visceral white fat at single-cell resolution across a range of body weight. We identify subpopulations of adipocytes, adipose stem and progenitor cells, vascular and immune cells and demonstrate commonalities and differences across species and dietary conditions. We link specific cell types to increased risk of metabolic disease and provide an initial blueprint for a comprehensive set of interactions between individual cell types in the adipose niche in leanness and obesity. These data comprise an extensive resource for the exploration of genes, traits and cell types in the function of white adipose tissue across species, depots and nutritional conditions. The top 3 key questions that this resource can answer: 1. How specific is my gene of interest to a particular cell type in adipose tissue? 2. Is the gene/pathway that I am studying in mouse adipose tissue also present in human adipose tissue (and is it regulated similarly in low vs high body weight)? 3. What are the changes in gene expression in a specific cell type at low vs high body weight? Resource link: https://singlecell.broadinstitute.org/single_cell/study/SCP1376/a-single-cell-atlas-of-human-and-mouse-white-adipose-tissue#study-summary Upcoming webinars schedule: https://dknet.org/about/webinar

1. A single cell atlas of human and mouse white adipose tissue
Margo Emont
dkNET Webinar Series
November 17, 2023

2. Why adipose tissue?
Rosen & Spiegelman, Cell 2014

3. Adipose tissue is a heterogeneous organ
Adipocytes:
White adipocytes: LEP+
Thermogenic adipocytes: UCP1+
Subcutaneous adipocytes: TBX15+
Visceral adipocytes: ITLN1+
Lean adipocytes: CFD+
Obese adipocytes: MRC2+
Adipose stem and progenitor
cells:
Adipose stem cells: DPP4+, PI16+
Committed preadipocytes:
ICAM1+, FABP4+
Aregs: CD142+, FMO+
Immune cells:
Macrophages:
MAFB+
Monocytes:
CYBB+
Dendritic cells:
FLT3+
Mast cells:
CPA3+
Neutrophils:
CSF3R+
B cells: MS4A1+
NK cells:
KLRD1+
T cells: IL7R+
Vascular cells:
Blood endothelial cells (arteriole,
venule, stalk): PECAM1+
Lymphatic endothelial cells:
PROX1+
Pericytes: STEAP4+
Smooth muscle cells: MYOCD+
Emont & Rosen, Curr Opin Genet Dev 2023

4. Adipose tissue remodels during obesity
Lean, insulin sensitive Obese, insulin resistant
Rosen & Spiegelman, Cell 2014

5. Human adipose single cell
Emont et. al. Nature 2022

6. Human adipose single cell
Emont et. al. Nature 2022

7. Human adipose single cell
Emont et. al. Nature 2022
Subcutaneous
Visceral

8. Single nuclear sequencing captures the expected cell
types in adipose tissue
166,149 total cells: 28,465 whole cell, 137,684 single nucleus

9. Single nuclear sequencing captures the expected cell
types in adipose tissue
166,149 total cells: 28,465 whole cell, 137,684 single nucleus

10. Single nuclear sequencing captures the expected cell
types in adipose tissue
166,149 total cells: 28,465 whole cell, 137,684 single nucleus

11. Some cell populations are affected by depot and/or
BMI

12. Mouse adipose single cell also finds expected cell
types

13. Mouse adipose single cell also finds expected cell
types
197,721 total cells

14. Mouse adipose single cell also finds expected cell
types
197,721 total cells

15. Some mouse cell populations are also affected by
depot

16. Comparison between human and mouse populations
Mouse Populations Human Populations

17. Human and mouse vascular cells
22,734 human vascular cells, 7,632 mouse vascular cells

18. Human and mouse mesothelial cells
30,482 human mesothelial cells, 14,947 mouse mesothelial cells

19. 34,268 human immune cells, 70,547 mouse immune cells
Human and mouse immune cells

20. 52,482 human ASPCs, 51,227 mouse ASPCs
Human and mouse adipose stem and progenitor
cells

21. Mapping mouse ASPC populations to other studies

22. Human and mouse adipocytes
Human Adipocytes Mouse Adipocytes
25,817 human adipocytes, 39,934 mouse adipocytes

23. Exploring the adipose atlas online

24. Exploring the adipose atlas online

25. Exploring the adipose atlas online

26. Exploring the adipose atlas online

27. Exploring the adipose atlas online

28. Exploring the adipose atlas online

29. Exploring the adipose atlas online

30. Exploring the adipose atlas online

31. Exploring the adipose atlas online

32. Exploring the adipose atlas online

33. Exploring the adipose atlas online

34. Exploring the adipose atlas online

35. Exploring the adipose atlas online

36. Exploring the adipose atlas online

37. Downloading adipose atlas data

38. Downloading adipose atlas data

39. Mapping your adipose data to adipose reference
data

40. Mapping your adipose data to adipose reference
data

41. Human Cell Atlas Adipose Network
https://forms.gle/eEjmiXuzPQorVtY49
Sign up for the adipose network!

42. Adipose network integration efforts
Human data
• Collect through Human Cell Atlas to avoid data
privacy issues
• Integration and comparison led by HCA
Adipose Network
Mouse data
• Either collect raw data or send pipeline for all
interested groups to run on their data
• Integration and comparison led by HCA
Adipose Network
Compare mouse and human adipose cell types

43. Adipose Network White paper
Form
committees
Adipose
network virtual
meeting
Prepare and
submit
manuscript
• Consensus cell type
annotations
• Consensus depot naming and
framework for reporting
specific location samples
were taken from
• Best practices for sample
collection and processing
• Best practices for
bioinformatic analyses
• Planned for early 2024
• Presentations on research
from the network
• Presentations from
committees about progress
that has been made
• Presentations from HCA
representatives about best
practices gleaned from
broader integration efforts

44. Human Cell Atlas Adipose Network
https://forms.gle/eEjmiXuzPQorVtY49
Sign up for the adipose network!

45. Future plans
• Integrate available human and
mouse adipose datasets to
create a consensus atlas
• Sequence other adipose depots
such as epicardial and bone
marrow fat and compare to
existing datasets
• Create an adipose tissue
knowledge portal where users
can explore existing adipose
tissue -omics datasets

46. Future plans
• Integrate available human and
mouse adipose datasets to
create a consensus atlas
• Sequence other adipose
depots such as epicardial and
bone marrow fat and compare
to existing datasets
• Create an adipose tissue
knowledge portal where users
can explore existing adipose
tissue -omics datasets

47. Future plans
• Integrate available human and
mouse adipose datasets to
create a consensus atlas
• Sequence other adipose depots
such as epicardial and bone
marrow fat and compare to
existing datasets
• Create an adipose tissue
knowledge portal where users
can explore existing adipose
tissue -omics datasets

48. Acknowledgements
Evan Rosen
Linus Tsai
Christopher Jacobs
Adam Essene
Danielle Tenen
Deepti Pant
Greg Westcott
Anton Gulko
Molly Veregge
Harini Srinivasan
Amrita Kar
Zach Kramer
Collaborators:
Orr Ashenberg
Aviv Regev
Saverio Cinti
Antonio Giordano
Georgia Colleluori
Angelica Di Vincenzo
Melina Claussnitzer
Hesam Dashti
Adam Stefek
Elizabeth McGonagle
Sophie Strobel
Samantha Laber
Amit Khera
Saaket Agrawal
Tune Pers
Anja Jørgensen
Anita Courcoulas
William Gourash
Randy Seeley
Nadia Bozadjieva
Explore the data!
RC2DK116691
K01DK134806
Join the adipose network!