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Oncology

Biospace Lab contributes to the great efforts devoted to cancer research, which represents a major concern for human health.

Bar, I. et al. Silencing of casein kinase 1 delta reduces migration and metastasis of
triple negative breast cancer cells
. Oncotarget (2018)


Non-targeted (control)
The primary tumor grows over time in triple negative breast cancer xenografts


Silencing of CSNK1D
CSNK1D knock-down slows down tumor development

CSNK1D knock-down prevents metastasis formation, while lung metastases develop in shNT triple negative breast cancer xenografts

The highly dynamic range of the camera
allows to follow-up the primary tumor
as well as to detect lung metastases.

Bar, I. et al. Silencing of casein kinase 1 delta reduces migration and metastasis of
triple negative breast cancer cells
. Oncotarget (2018)

1st line: Non-targeted (control)
The primary tumor grows over time in triple negative breast cancer xenografts

2nd line Silencing of CSNK1D
CSNK1D knock-down slows down tumor development

CSNK1D knock-down prevents metastasis formation, while lung metastases develop in shNT triple negative breast cancer xenografts

The highly dynamic range of the camera allows to follow-up the primary tumor as well as to detect lung metastases.

Jarry, U. et al. Orthotopic model of lung cancer: isolation of bone micro-metastases
after tumor escape from Osimertinib treatment
. BMC Cancer (2021)

Orthotopic model of lung cancer:
– lung primary tumor nesting
– metastases dissemination after Osimertinib treatment

Bone micro-metastases were easily detected using bioluminescence, allowing isolation of tumor cells otherwise undetectable by macroscopic observation.

Germanova, D. et al. Myeloid tumor necrosis factor and heme oxygenase‐1 regulate the progression of colorectal liver metastases during hepatic ischemia‐reperfusion. International Journal of Cancer (2021)

Bioluminescence imaging. CCD camera image capture of C57BL/6 mice inoculated 14 days before with luciferase-MC-38 cells (T group) and undergoing liver IR 12 days before (T + IR group)
Serial liver emitted photon quantification at 9 and 14 days postluciferase-MC-38 inoculation in C57BL/6 mice (*P < .05, and ***P < .001 two-tailed nonparametric Mann-Whitney test). IR, ischemia-reperfusion

Bogeas, A. et al. Changes in chromatin state reveal ARNT2 at a node of a tumorigenic transcription factor signature driving glioblastoma cell aggressiveness. Acta Neuropathologica (2018)

 shCTL (control)
Mice with
orthotopic xenograft of glioblastoma
stem-like cells (left) almost inevitabely develop a tumor 

Silencing of ARNT2
ARNT2 knock-down (right) strikingly reduce tumor incidence in mice grafted with glioblastoma
stem-like cells

Longitudinal analysis of bioluminescence allows to reveal tumor incidence and the timing of its occurrence.

 

Bogeas, A. et al. Changes in chromatin state reveal ARNT2 at a node of a tumorigenic transcription factor signature driving glioblastoma cell aggressiveness. Acta Neuropathologica (2018)

 shCTL (control)
Mice with orthotopic xenograft of glioblastoma stem-like cells (left) almost inevitabely develop a tumor 

shARNT2: Silencing of ARNT2
ARNT2 knock-down (right) strikingly reduce tumor incidence in mice grafted with glioblastoma stem-like cells

Longitudinal analysis of bioluminescence allows to reveal tumor incidence and the timing of its occurrence.

Arfaoui, A. et al. A genome‐wide RNAi screen reveals essential therapeutic targets of breast cancer stem cells. EMBO Molecular Medicine (2019)

Untreated (control)
Primary breast cancer xenografts engender liver and lung metastasis formation in untreated mice
Salinomycin treated
Salinomycin alone does not significantly impair metastasis development
JQ1 treated
JQ1 alone does not significantly impair metastasis development

Salinomycin/ JQ1 treated
The salinomycin/JQ1 combination is efficent for significant reduction of metastasis formation

Treatment efficacy on metastasis development is quantified in two primary human breast cancer xenograft models (CRCM434 and 404), generated from patients with chemo-naïve triple-negative breast tumors.

Drug combination assay and ex vivo imaging display the synergistic interaction of salinomycin/JQ1 association to limit breast cancer metastatic development.

Arfaoui, A. et al. A genome‐wide RNAi screen reveals essential therapeutic targets of breast cancer stem cells. EMBO Molecular Medicine (2019)

CTRL: Untreated (control)
Primary breast cancer xenografts engender liver and lung metastasis formation in untreated mice
SAL: Salinomycin treated
Salinomycin alone does not significantly impair metastasis development
JQ1: JQ1 treated
JQ1 alone does not significantly impair metastasis development

SAL+JQ1: Salinomycin/ JQ1 treated
The salinomycin/JQ1 combination is efficent for significant reduction of metastasis formation

Treatment efficacy on metastasis development is quantified in two primary human breast cancer xenograft models (CRCM434 and 404), generated from patients with chemo-naïve triple-negative breast tumors.

Drug combination assay and ex vivo imaging display the synergistic interaction of salinomycin/JQ1 association to limit breast cancer metastatic development.

Neurology

The central nervous system is a complex and fascinating structure that remains yet complicated to image. Biospace Lab supports neurobiologists in this challenge

Guglielmetti, C. Prael, J. et al. Multimodal imaging of subventricular zone neural stem/progenitor cells in the cuprizone mouse model reveals increased neurogenic potential for the olfactory bulb pathway, but no contribution to remyelination of the corpus callosum. NeuroImage (2014)


Untreated (control)

In situ labeled NPCs of the subventricular zone (SVZ) migrate towards the olfactory bulb (OB)

 

Multiple sclerosis model
Significantly increased migration of NPCs from the SVZ to the OB is evidenced in cuprizone-treated mice

Guglielmetti1

Longitudinal in vivo tracking allows
monitoring neural progenitor cells (NPCs)
within distinct brain regions.

Increased proliferation and significant alteration of NPC migration from the SVZ to the OB has been highlighted in a murine model of multiple sclerosis, induced by cuprizone treatement.

Guglielmetti, C. Prael, J. et al. Multimodal imaging of subventricular zone neural stem/progenitor cells in the cuprizone mouse model reveals increased neurogenic potential for the olfactory bulb pathway, but no contribution to remyelination of the corpus callosum. NeuroImage (2014)

Guglielmetti1

Untreated (control)
In situ labeled NPCs of the subventricular zone (SVZ) migrate towards the olfactory bulb (OB)

Cuprizone: Multiple sclerosis model
Significantly increased migration of NPCs from the SVZ to the OB is evidenced in cuprizone-treated mice

              Longitudinal in vivo tracking
              allows monitoring neural
              progenitor cells (NPCs)
              within distinct brain regions.

Increased proliferation and significant alteration of NPC migration from the SVZ to the OB has been highlighted in a murine model of multiple sclerosis, induced by cuprizone treatement.

Gakuba, C. et al. General anesthesia inhibits the activity of the “glymphatic system”. Theranostics (2018)

 

Awake
The indocyanine green fluorescent tracer injected into the cisterna magna (orange) diffuses through the brain and is readily detectable in the forebrain (red)


Anesthetized (isoflurane)
The intracisternally injected indocyanine green remains near the injection site in the anesthetized mice

Gakuba

As soon as 30 min after intracisternal injection, the amount of indocyanine green in the forebrain of awake mice is significantly higher with respect to anesthetized mice.

Longitudinal in vivo near infrared imaging of freely moving mice revealed that cerebrospinal fluid flow circulation is mainly active during wakefulness
and significantly impaired during
general anesthesia.

Gakuba, C. et al. General anesthesia inhibits the activity of the “glymphatic system”. Theranostics (2018)

Gakuba mobile

Awake
The indocyanine green fluorescent tracer injected into the cisterna magna (orange) diffuses through the brain and is readily detectable in the forebrain (red)

Anesthetized (isoflurane)
The intracisternally injected indocyanine green remains near the injection site in the anesthetized mice

As soon as 30 min after intracisternal injection, the amount of indocyanine green in the forebrain of awake mice is significantly higher with respect to anesthetized mice.

Longitudinal in vivo near infrared imaging of freely moving mice revealed that cerebrospinal fluid flow circulation is mainly active during wakefulness and significantly impaired during general anesthesia.

Rogers, K. L. et al. Non-Invasive In Vivo Imaging of Calcium Signaling in Mice. PLoS ONE (2007)

Seizure induced by kainate administration in non-anesthetized mice, provokes highly synchronized, bi-lateral Ca2+ transients.

This increased Ca2+ activity is measured using GFP-aequorin as a bioluminescent Ca2+-reporter.

Real-time bioluminescent signal acquisition allows to detects
rapid Ca2+ transients.

A marked increase in Ca2+ activity occurred approximately 15 minutes after drug administration.
This phenomenon is evidenced as oscillations in which Ca2+ spikes only last a few seconds.

Stem Cell Biology

Stem cells are essential actors in embryonic development and tissue homeostasis and renewal. Biospace Lab proposes versatile solutions that offer decisive advantages for investigating and exploiting their exclusive potential.

Tennstaedt, A. et al. Human neural stem cell intracerebral grafts show spontaneous early neuronal differentiation after several weeks. Biomaterials (2015)


Early neuronal reporter

Newly born neuron signal increases within the first month after NSC implantation

 

Constitutively expressed reporter (control)
The signal remains stable over time

 

 

 

Mature neuronal reporter
Ongoing maturation of neurons and synaptogenesis is detectable from 3 months post-implantation

Tennstaedt, A. et al. Human neural stem cell intracerebral grafts show spontaneous early neuronal differentiation after several weeks. Biomaterials (2015)

1st line: Early neuronal reporter
Newly born neuron signal increases within the first month after NSC implantation

2nd line: Constitutively expressed reporter (control)
The signal remains stable over time

3rd line: Mature neuronal reporter
Ongoing maturation of neurons and synaptogenesis is detectable from 3 months post-implantation

Virology / infectiology

Pavlidis, I. et al. Cervical epithelial damage promotes Ureaplasma parvum ascending infection, intrauterine inflammation and preterm birth induction in mice. Nature Communications (2020)

Controls
No signal is detectable in untreated (PBS) or in N-9 treated mice when not subjected to infection (USM)

Untreated
Low bacterial load is observable in untreated mice infected with a luciferase-expressing Ureaplasma parvum strain (UP)

N-9 induced cervical damage
Intravaginal exposure to N-9 facilitates intrauterine UP infection 


Bioluminescent bacteria emanate
high-intensity signals with the expected
shape and location of the uterus, showing bulges representing gestating foetus

In vivo imaging allows to visualize the ascending
Ureaplasma parvum infection in pregnancy,
demonstrating the crucial role of the cervical epithelium
as a barrier against ascending infection and preterm birth.

Pavlidis, I. et al. Cervical epithelial damage promotes Ureaplasma parvum ascending infection, intrauterine inflammation and preterm birth induction in mice. Nature Communications (2020)

Controls
No signal is detectable in untreated (PBS) or in N-9 treated mice when not subjected to infection (USM)

Untreated
Low bacterial load is observable in untreated mice infected with a luciferase-expressing Ureaplasma parvum strain (UP)

N-9 induced cervical damage
Intravaginal exposure to N-9 facilitates intrauterine UP infection

Bioluminescent bacteria emanate
high-intensity signals with the expected shape and location of the uterus, showing bulges representing gestating foetuses

In vivo imaging allows to visualize the ascending Ureaplasma parvum infection in pregnancy, demonstrating the crucial role of the cervical epithelium as a barrier againstascending infection and preterm birth.

Raaben, M. et al. Non-invasive imaging of mouse hepatitis coronavirus infection reveals determinants of viral replication and spread in vivo. Cellular Microbiology (2009)

Mice with different genetic background have been intranasally inoculated with
lucifesare-expressing mouse hepatitis virus (MHV)

BALB/c mouse strain
BALB/c mice display the highest luminescent signal

C57BL/6 mouse strain
C57BL/6 mice have an intermediate phenotype compared to the other genetic backgrounds

129SvEv mouse strain
129SvEv mice are significantly less susceptible to MHV infection

Non-invasive whole-body imaging of viral infection
in living animals allows to monitor its spatial
and temporal progression in real-time.

Follow-up of viral replication can be performed
in parallel for different animals, here unveiling
the variable susceptibility of various
inbred mouse strains to MHV infections.

Others

Virtually any beta emitting radioisotopes with sufficient energy will generate Cerenkov luminescence in vivo , including those for which no current in vivo imaging solutions exist; such as 32P, 90Y and 90Sr.

2.5 μCi of 90Y radiotracer concentrated in the pancreas of a mouse—imaged with PhotonIMAGER         

Cerenkov luminescence with the 4-View module of the PhotonIMAGER™

Images of a mouse from 4 different angles 4h after the injection of 50 μCi 32P. Cerenkov luminescence signal appears in subcutaneous tumor — with a high pass (530nm) filter.

Weinmann, J. et al. Identification of a myotropic AAV by massively parallel in vivo evaluation of barcoded capsid variants. Nature Communications (2020)

Pharmacology / biodistribution

Zinnhardt, B. et al. Molecular imaging of immune cell dynamics during de- and remyelination in the cuprizone model of multiple sclerosis by [18F]DPA-714 PET and MRI. Theranostics (2019)

Comparison of in vivo PET data with ex vivo autoradiography. (A) Ex vivo autoradiography confirms the in vivo spatial distribution of [18F]DPA-714 (n = 1-2).

Kryza, D. et al. Ex vivo and in vivo imaging and biodistribution of aptamers targeting the human Matrix MetalloProtease-9 in melanomas. PLoS ONE (2016)

Comparison of the results obtained by radiolabeling of representative tumor tissue sections with 111In-DOTA-F3B aptamer (left image) and 111In-DOTA-control sequence (right image).The difference of activity seems to increase in a tumor grade-dependent manner. (A) Lentigo malignant melanoma, (B) Nodular melanoma, (C) Mostly metastatic node.

Alitalo, O., Rantamäki, T. & Huhtala, T. Digital autoradiography for efficient functional imaging without anesthesia in experimental animals: Reversing phencyclidine-induced functional alterations using clozapine. Progress in Neuro-Psychopharmacology and Biological Psychiatry (2020)

With the advances in direct particle-counting systems, the new digital ARG devices have surpassed the conventional film and phosphor screen-based ARG in linearity, dynamic range, and sensitivity, reducing the acquisition times from months to just few hours.”

 

An example of qualitative results acquired from control group with BetaImager TRacer using 3H-labeled 2-deoxy-d-glucose as a tracer. The acquisition was performed for 6 h on a full-field zoom setting (250 × 200 mm)”

Alitalo, O., Rantamäki, T. & Huhtala, T. Digital autoradiography for efficient functional imaging without anesthesia in experimental animals: Reversing phencyclidine-induced functional alterations using clozapine. Progress in Neuro-Psychopharmacology and Biological Psychiatry (2020)

With the advances in direct particle-counting systems, the new digital ARG devices have surpassed the conventional film and phosphor screen-based ARG in linearity, dynamic range, and sensitivity, reducing the acquisition times from months to just few hours.”

 

An example of qualitative results acquired from control group with BetaImager TRacer using 3H-labeled 2-deoxy-d-glucose as a tracer. The acquisition was performed for 6 h on a full-field zoom setting (250 × 200 mm)”