Persistent confined migration confers permanent nuclear and functional changes in migrating cells
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Abstract
Nuclear deformability plays a critical role in cell migration. During this process, the remodeling of internal components of the nucleus has a direct impact on DNA damage and cell behavior; however, how persistent migration promotes nuclear changes leading to phenotypical and functional consequences remains poorly understood. Here, we described that the persistent migration through physical barriers was sufficient to promote permanent modifications in migratory-altered cells. We found that lamin B1 altered its localization, concomitant with morphological and transcriptional changes. Migratory-altered cells showed alterations in cellular functions such as DNA repair and cell migration. We applied biochemical and biophysical approaches to identify that confined conditions altered the biomechanical response of the nucleus. Mechanistically, we determined that actin dynamics controlled the redistribution of lamin, and the basal levels of DNA damage in migratory-altered cells. Our observations reveal a novel role for confined cell conditions in consistent nuclear and genomic alterations that might handle the genetic instability and cellular heterogeneity in aging diseases and cancer.
Highlights
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Persistent confined migration promotes permanent mophological changes.
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Lamin B1 is redistribution in the nucleus of migratory altered cells.
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Migratory-altered cells exhibit transcriptional and functional changes related to cell migration and survival.
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Actin polymerization controls nuclear changes induced by cell migration.
Article activity feed
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then culturing them in the absence of further mechanical stimuli
Very cool process! Have you taken the MA cells through several passages an they continue to retain this phenotype?
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urkat and CCRF-CEM
I really appreciate the consistency of phenotype across both cell types! I'm curious if a similar phenotype would occur in non-cancerous cells as well!
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o investigate the effect of persistent migration on the nuclear changes of MA cells, we determined their lamin B1 distribution and found that MA cells showed an aberrant distribution of lamin B1
Were the MA cells in suspended culture for at least 24 hours (like the recovered ORM cells)?
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by allowing cells to migrate three–rounds across rigid filter membranes
Interesting! Is this something that would be happening in living tissue? I'd be curious to hear if y'all tried different membrane materials or numbers of passages!
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1.8-fold change in 639
Wow! That's a big change in 3 days!!! I'd be interested in seeing images of more than just the nucleus. Are there cell shape changes in the MA cells compared to the control?
Also, I'd be interested to see a comparison between control, ORM, and MA cells! Since the ORM cells can revert back to "normal" cells, it'd be interesting to compare the expression in revertable and non-revertable cells to get more insight into the genes responsible for this reversion!
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expanded
Very cool! I'm guessing "expanded" means the cells were split/passaged, right? It could be helpful if this is clearly defined since expand could be interpreted as expansion microscopy preparation which would lead to larger nuclei. It is super cool that these phenotypes persist!
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1 (Figures 2F, 2G, S2F, S2G, and Movies S3, S4)
The Y-axes aren't consistent here so its hard to tell, but it seems like there is much stronger whole nucleus lamin signal in MA cells compared to control. Is this correct? Have y'all quantified the whole cell levels of lamin in MA & control cells?
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low levels of γH2AX (<2 foci per nucleus),
I'm curious why <2 foci is the measurement here. It seems whole cell intensity or a full # of foci count would show greater variation between the conditions (based solely on the representative images). Sorry if this is a common assay that is frequently done.
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To gain insight into how persistent migration might affect cell viability, we treated control and MA cells with methotrexate, a DNA biosynthesis inhibitor used in clinics against multiple human pathologies [26]. Interestingly, we found that MA cells showed higher resistance to apoptosis than control cells (Fig. 4E, 4F).
That is wild! Have y'all looked at other inducers of apoptosis? Is it possible that the methotrexate just isn't getting into the nucleus as much (since MA cells seem to have so much more lamin surrounding & throughout the nucleus).
Oh! I see you used staurosporine which is listed as an apoptosis inducer! Did y'all notice any changes in survival rate when treated with Staurosporine?
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These results indicate that constrained migration alters the mechanical behavior of the nucleus, which might have functional implications on the capacity of MA cells to infiltrate tissues.
Interesting! So the MA cells will migrate more than control cells through collagen, but are less capable through animal tissue! Also it is incredibly strange that the Jurkat MAs are found significantly less in BM and Liver (but not Spleen), but the CCRF-CEM MA cells are significantly less in Spleen an Liver (but not BM), right!??!
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These results indicate that constrained migration alters the mechanical behavior of the nucleus, which might have functional implications on the capacity of MA cells to infiltrate tissues.
Interesting! So the MA cells will migrate more than control cells through collagen, but are less capable through animal tissue! Also it is incredibly strange that the Jurkat MAs are found significantly less in BM and Liver (but not Spleen), but the CCRF-CEM MA cells are significantly less in Spleen an Liver (but not BM), right!??!
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low levels of γH2AX (<2 foci per nucleus),
I'm curious why <2 foci is the measurement here. It seems whole cell intensity or a full # of foci count would show greater variation between the conditions (based solely on the representative images). Sorry if this is a common assay that is frequently done.
-
by allowing cells to migrate three–rounds across rigid filter membranes
Interesting! Is this something that would be happening in living tissue? I'd be curious to hear if y'all tried different membrane materials or numbers of passages!
-
then culturing them in the absence of further mechanical stimuli
Very cool process! Have you taken the MA cells through several passages an they continue to retain this phenotype?
-
To gain insight into how persistent migration might affect cell viability, we treated control and MA cells with methotrexate, a DNA biosynthesis inhibitor used in clinics against multiple human pathologies [26]. Interestingly, we found that MA cells showed higher resistance to apoptosis than control cells (Fig. 4E, 4F).
That is wild! Have y'all looked at other inducers of apoptosis? Is it possible that the methotrexate just isn't getting into the nucleus as much (since MA cells seem to have so much more lamin surrounding & throughout the nucleus).
Oh! I see you used staurosporine which is listed as an apoptosis inducer! Did y'all notice any changes in survival rate when treated with Staurosporine?
-
1.8-fold change in 639
Wow! That's a big change in 3 days!!! I'd be interested in seeing images of more than just the nucleus. Are there cell shape changes in the MA cells compared to the control?
Also, I'd be interested to see a comparison between control, ORM, and MA cells! Since the ORM cells can revert back to "normal" cells, it'd be interesting to compare the expression in revertable and non-revertable cells to get more insight into the genes responsible for this reversion!
-
1 (Figures 2F, 2G, S2F, S2G, and Movies S3, S4)
The Y-axes aren't consistent here so its hard to tell, but it seems like there is much stronger whole nucleus lamin signal in MA cells compared to control. Is this correct? Have y'all quantified the whole cell levels of lamin in MA & control cells?
-
o investigate the effect of persistent migration on the nuclear changes of MA cells, we determined their lamin B1 distribution and found that MA cells showed an aberrant distribution of lamin B1
Were the MA cells in suspended culture for at least 24 hours (like the recovered ORM cells)?
-
expanded
Very cool! I'm guessing "expanded" means the cells were split/passaged, right? It could be helpful if this is clearly defined since expand could be interpreted as expansion microscopy preparation which would lead to larger nuclei. It is super cool that these phenotypes persist!
-
urkat and CCRF-CEM
I really appreciate the consistency of phenotype across both cell types! I'm curious if a similar phenotype would occur in non-cancerous cells as well!
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