- Glioblastoma multiforme (GBM) is a highly aggressive brain tumor whose migration patterns are not well understood. - Bioluminescent imaging was used to monitor GBM tumors implanted in mice in vivo to identify reproducible migration patterns. - Distinct migration patterns were observed, including migration into the olfactory bulb and subventricular zone, as well as more common spinal metastases than previously thought. Migration was also found to surround the exterior of the cerebellum.

1. ● GBM is a highly aggressive and invasive
primary brain tumor whose migration patterns
are not yet well understood 1
.
● We used bioluminescent imaging (BLI) to
monitor tumors in vivo.
● Goal - find a reproducible pattern of migration
that can be targeted by future drug therapies.
Acknowledgments
We want to thank Dr. Sanjiv S. Gambhir and Google for
supporting the internship program. We also want to thank
our mentors, Dr. Edwin Chang and Dr. Christoph Pohling,
for their guidance and support.
Conclusions
● Distinct and reproducible migration pattern
○ Observed with multiple patient-derived
cell lines
○ Migration into olfactory bulb and
possible subventricular zone
○ Spinal metastases are more common than
previously thought because even when
spine metastasis was not visible in the
mouse, one can see it in the spine section
○ GBM migration only surround the
exterior of cerebellum and did not enter
the cerebellum area.
Kathryn Li*, Alex Serafini*, Tara Thakurta*, Xiaofan Wu*, Edwin Chang, and Christoph Pohling
3155 Porter Drive, Palo Alto, CA 94304, USA
* A.S., T.T., K.L., and X.W. contributed equally to this project
Mersereau, J., Levy, N., Staub, R., Baggett, S., Zogric, T., Chow, S., . . . Bjeldanes, L. (2008). Liquiritigenin is a plant-derived highly selective estrogen receptor β agonist. Molecular and Cellular Endocrinology, 49
1. H. Adams, J. Avendano, S.M. Raza, Z.L. Gokaslan, G.I. Jallo, A. Quinones-Hinojosa. Prognostic Factors and Survival in Primary Malignan Astrocytomas of the Spinal Cord. Spine Volume 37, Number 12: E727-E735, 2007.
2. J.S. Guillamo, F. Lisovoski, C. Christov, C. Le Guerinel, G.L. Defer, M. Peschanski and T. Lefrancois. Migration pathways of human glioblastoma cells xenografted into the immunosuppressed rat brain. Journal of Neuro-Oncology 52: 205-215, 2001.
3. J. Knierim. “Chapter 5: Cerebellum.” Neuroscience Online. 1997. <http://neuroscience.uth.tmc.edu/s3/chapter05.html>.
4. K. Onda, R. Tanaka, H. Takahashi, N. Takeda, F. Ikuta. Cerebral Glioblastoma with Cerebrospinal Fluid Dissemination: A Clinicopathological Study of 14 Cases Examined by Complete Autopsy. Neurosurgery Volume 25, Issue 4: 533-540.
5. H. Sontheimer. S. Watkins. Unique biology of gliomas: challenges and opportunities. Trends in Neurosciences Volume 35, Issue 9: 546-556, 2012.
Future Directions
● Investigate the major mechanisms of
migration: CSF, white matter, or vasculature
tracking?
● Experiment with different sites of
implantation
● FACS separation of GBM cells by forward
side-scatter in order to study effects of size
heterogeneity with regards to migration
patterns
● Research anatomically selective drug
treatments
Initial BLI slice imaging of drug effects on
GBM migration, with 3D reconstruction for
more accurate drug targeting pending.
3D Reconstruction of Human Glioma in Mice
2015 Internship program supported by:
Introduction
Materials and Methods
Results Conclusions
Literature cited
Future Directions
Acknowledgments
Doublet PatternSinglet Pattern
Section brain
and analyze w/
BLI
BLI monitoring of
growing tumors
Tumor
implantation
● pcGBM2 GFP/luc cells were implanted in
nude mice.
● Mice were imaged weekly to monitor tumor
growth.
● Sections were obtained from harvested
mouse brains and imaged with BLI to identify
migration patterns. A 3D rendering of a brain with a GBM39 doublet tumor.
The olfactory bulb appears to be a niche for GBM.
Correlating Migratory Pathways with Anatomical Locations
● GBM appears to travel via specific anatomical structures. In Figure 2,
GBM seems to track along the white matter.
● The images also show an accumulation of tumor cells between the
cerebellum and the cerebrum, suggesting migration through
cerebrospinal fluid (CSF).
● The isolated BLI signals throughout the brain shown in Figure 1 are
associated with GBM accumulation around vasculature.
● Future investigation is necessary to identify the cause of migration along
these particular anatomical pathways.
A 3D rendering of a brain implanted with pcGBM2. The
model displays the migration pathway that we identified.
Spinal Metastasis
8% 67% 25%
3D Reconstructions
After collecting approximately 40
slices (25 microns in width) from
two patient derived cell lines,
ImageJ was used to render a 3D
model. The BLI signal correlates to
the presence of GBM xenograft,
allowing for observation of both
migration pathways and niches
favorable for tumor formation.
pcGBM2 cell line LN229 cell line U87 cell line
a
fe
dc
b
Figure 2:Individual Slices
Figure 1:Initial Studies
Figure 3:3D Reconstructions
a b

2. ● GBM is a highly aggressive and invasive
primary brain tumor whose migration patterns
are not yet well understood1
.
● We used bioluminescent imaging (BLI) to
monitor tumors in vivo.
● Goal - find a reproducible pattern of migration
that can be targeted by future drug therapies.
We want to thank Dr. Sanjiv S. Gambhir and Google for
supporting the internship program. We also want to thank
our mentors, Dr. Edwin Chang and Dr. Christoph Pohling,
for their guidance and support.
Conclusions
● Distinct and reproducible migration pattern
○ Observed with multiple patient-derived
cell lines
○ Migration into olfactory bulb and
possible subventricular zone
○ Spinal metastases are more common than
previously thought because even when
spine metastasis was not visible in the
mouse, one can see it in the spine section
○ GBM migration only surround the
exterior of cerebellum and did not enter
the cerebellum area.
Kathryn Li*, Alex Serafini*, Tara Thakurta*, Xiaofan Wu*, Edwin Chang, and Christoph Pohling
3155 Porter Drive, Palo Alto, CA 94304, USA
* A.S., T.T., K.L., and X.W. contributed equally to this project
Mersereau, J., Levy, N., Staub, R., Baggett, S., Zogric, T., Chow, S., . . . Bjeldanes, L. (2008). Liquiritigenin is a plant-derived highly selective estrogen receptor β agonist. Molecular and Cellular Endocrinology, 49
1. H. Adams, J. Avendano, S.M. Raza, Z.L. Gokaslan, G.I. Jallo, A. Quinones-Hinojosa. Prognostic Factors and Survival in Primary Malignan Astrocytomas of the Spinal Cord. Spine Volume 37, Number 12: E727-E735, 2007.
2. J.S. Guillamo, F. Lisovoski, C. Christov, C. Le Guerinel, G.L. Defer, M. Peschanski and T. Lefrancois. Migration pathways of human glioblastoma cells xenografted into the immunosuppressed rat brain. Journal of Neuro-Oncology 52: 205-215, 2001.
3. J. Knierim. “Chapter 5: Cerebellum.” Neuroscience Online. 1997. <http://neuroscience.uth.tmc.edu/s3/chapter05.html>.
4. K. Onda, R. Tanaka, H. Takahashi, N. Takeda, F. Ikuta. Cerebral Glioblastoma with Cerebrospinal Fluid Dissemination: A Clinicopathological Study of 14 Cases Examined by Complete Autopsy. Neurosurgery Volume 25, Issue 4: 533-540.
5. H. Sontheimer. S. Watkins. Unique biology of gliomas: challenges and opportunities. Trends in Neurosciences Volume 35, Issue 9: 546-556, 2012.
Future Directions
● Investigate the major mechanisms of
migration: CSF, white matter, or vasculature
tracking?
● Experiment with different sites of
implantation
● FACS separation of GBM cells by forward
side-scatter in order to study effects of size
heterogeneity with regards to migration
patterns
● Research anatomically selective drug
treatments
Initial BLI slice imaging of drug effects on
GBM migration, with 3D reconstruction for
more accurate drug targeting pending.
3D Reconstruction of Human Glioma in Mice
2015 Internship program supported by:
Introduction
Materials and Methods
Conclusions
Literature cited
Future Directions
Acknowledgments
Doublet PatternSinglet Pattern
Section brain
and analyze w/
BLI
BLI monitoring of
growing tumors
Tumor
implantation
● pcGBM2 GFP/luc cells were implanted in
nude mice.
● Mice were imaged weekly to monitor tumor
growth.
● Sections were obtained from harvested
mouse brains and imaged with BLI to identify
migration patterns. A 3D rendering of a brain with a GBM39 doublet tumor.
The olfactory bulb appears to be a niche for GBM.
Correlating Migratory Pathways with Anatomical Locations
● GBM appears to travel via specific anatomical structures. In Figure 2,
GBM seems to track along the white matter.
● The images also show an accumulation of tumor cells between the
cerebellum and the cerebrum, suggesting migration through
cerebrospinal fluid (CSF).
● The isolated BLI signals throughout the brain shown in Figure 1 are
associated with GBM accumulation around vasculature.
● Future investigation is necessary to identify the cause of migration along
these particular anatomical pathways.
A 3D rendering of a brain implanted with pcGBM2. The
model displays the migration pathway that we identified.
Spinal Metastasis
8% 67% 25%
3D Reconstructions
After collecting approximately 40
slices (25 microns in width) from
two patient derived cell lines,
ImageJ was used to render a 3D
model. The BLI signal correlates to
the presence of GBM xenograft,
allowing for observation of both
migration pathways and niches
favorable for tumor formation.
pcGBM2 cell line LN229 cell line U87 cell line
a
fe
dc
b
Figure 2:Individual Slices
Figure 1:Initial Studies
Figure 3:3D Reconstructions
a b
Results

3. ● GBM is a highly aggressive and invasive
primary brain tumor whose migration patterns
are not yet well understood 1
.
● We used bioluminescent imaging (BLI) to
monitor tumors in vivo.
● Goal - find a reproducible pattern of migration
that can be targeted by future drug therapies.
Acknowledgments
We want to thank Dr. Sanjiv S. Gambhir and Google for
supporting the internship program. We also want to thank
our mentors, Dr. Edwin Chang and Dr. Christoph Pohling,
for their guidance and support.
Conclusions
● Distinct and reproducible migration pattern
○ Observed with multiple patient-derived
cell lines
○ Migration into olfactory bulb and
possible subventricular zone
○ Spinal metastases are more common than
previously thought because even when
spine metastasis was not visible in the
mouse, one can see it in the spine section
○ GBM migration only surround the
exterior of cerebellum and did not enter
the cerebellum area.
Kathryn Li*, Alex Serafini*, Tara Thakurta*, Xiaofan Wu*, Edwin Chang, and Christoph Pohling
3155 Porter Drive, Palo Alto, CA 94304, USA
* A.S., T.T., K.L., and X.W. contributed equally to this project
Mersereau, J., Levy, N., Staub, R., Baggett, S., Zogric, T., Chow, S., . . . Bjeldanes, L. (2008). Liquiritigenin is a plant-derived highly selective estrogen receptor β agonist. Molecular and Cellular Endocrinology, 49
1. H. Adams, J. Avendano, S.M. Raza, Z.L. Gokaslan, G.I. Jallo, A. Quinones-Hinojosa. Prognostic Factors and Survival in Primary Malignan Astrocytomas of the Spinal Cord. Spine Volume 37, Number 12: E727-E735, 2007.
2. J.S. Guillamo, F. Lisovoski, C. Christov, C. Le Guerinel, G.L. Defer, M. Peschanski and T. Lefrancois. Migration pathways of human glioblastoma cells xenografted into the immunosuppressed rat brain. Journal of Neuro-Oncology 52: 205-215, 2001.
3. J. Knierim. “Chapter 5: Cerebellum.” Neuroscience Online. 1997. <http://neuroscience.uth.tmc.edu/s3/chapter05.html>.
4. K. Onda, R. Tanaka, H. Takahashi, N. Takeda, F. Ikuta. Cerebral Glioblastoma with Cerebrospinal Fluid Dissemination: A Clinicopathological Study of 14 Cases Examined by Complete Autopsy. Neurosurgery Volume 25, Issue 4: 533-540.
5. H. Sontheimer. S. Watkins. Unique biology of gliomas: challenges and opportunities. Trends in Neurosciences Volume 35, Issue 9: 546-556, 2012.
Future Directions
● Investigate the major mechanisms of
migration: CSF, white matter, or vasculature
tracking?
● Experiment with different sites of
implantation
● FACS separation of GBM cells by forward
side-scatter in order to study effects of size
heterogeneity with regards to migration
patterns
● Research anatomically selective drug
treatments
Initial BLI slice imaging of drug effects on
GBM migration, with 3D reconstruction for
more accurate drug targeting pending.
3D Reconstruction of Human Glioma in Mice
2015 Internship program supported by:
Introduction
Materials and Methods
Results Conclusions
Literature cited
Future Directions
Acknowledgments
Doublet PatternSinglet Pattern
Section brain
and analyze w/
BLI
BLI monitoring of
growing tumors
Tumor
implantation
● pcGBM2 GFP/luc cells were implanted in
nude mice.
● Mice were imaged weekly to monitor tumor
growth.
● Sections were obtained from harvested
mouse brains and imaged with BLI to identify
migration patterns. A 3D rendering of a brain with a GBM39 doublet tumor.
The olfactory bulb appears to be a niche for GBM.
Correlating Migratory Pathways with Anatomical Locations
● GBM appears to travel via specific anatomical structures. In Figure 2,
GBM seems to track along the white matter.
● The images also show an accumulation of tumor cells between the
cerebellum and the cerebrum, suggesting migration through
cerebrospinal fluid (CSF).
● The isolated BLI signals throughout the brain shown in Figure 1 are
associated with GBM accumulation around vasculature.
● Future investigation is necessary to identify the cause of migration along
these particular anatomical pathways.
A 3D rendering of a brain implanted with pcGBM2. The
model displays the migration pathway that we identified.
Spinal Metastasis
8% 67% 25%
3D Reconstructions
After collecting approximately 40
slices (25 microns in width) from
two patient derived cell lines,
ImageJ was used to render a 3D
model. The BLI signal correlates to
the presence of GBM xenograft,
allowing for observation of both
migration pathways and niches
favorable for tumor formation.
pcGBM2 cell line LN229 cell line U87 cell line
a
fe
dc
b
Figure 2:Individual Slices
Figure 1:Initial Studies
Figure 3:3D Reconstructions
a b