HAMLET kills human glioblastoma cells in brain xenografts by an apoptosis-like mechanism and prolongs survival

1.  HAMLET AGAINST
CANCER
 HAMLET kills
human
glioblastoma cells
in brain xenografts
by an apoptosis-
like mechanisms
and prolongs
survival…….

2. • HAMLET ( Human Alpha-lactalbumin Made LEthal
to Tumor cells) is a protein-lipid complex that induces
apoptosis-like death in tumor cells but leaves fully
unaffected.
• HAMLET limits the progression of human glioblastomas
in xenograft model & removes skin papillomas in
patients.
• This broad anti-tumor activity includes >40 different
lymphomas & carcinomas & apoptosis is independent of
p53 or bcl-2.

4.  HAMLET structure and function in vitro.
A. HAMLET is formed from native α-lactalbumin by removal of Ca2 and by addition of the C18:1,
9 cis fatty acid (13). The figure is based on the α-lactalbumin crystal structure (33).
B. sensitivity of tumor cell lines to HAMLET. LD50, concentration required to kill 50% of the cells
in 6/24 h.
C. dose response of glioma cell lines to HAMLET. Murine primary brain cell cultures were used as
controls.
D. resistance of non-transformed human astrocytes to HAMLET treatment (0.03 mM).
E. cellular trafficking of HAMLET in malignant glioma cells. HAMLET (red, top panels) binds to
the tumor cell surface, invades their cytoplasm, translocates to the perinuclear region,
and accumulates in the nuclei. Morphological changes include membrane blebbing, nuclear
condensation, vesicle formation, cell shrinkage, and formation of apoptotic bodies (light
transmission; bottom panels, magnification, 180). α-Lactalbumin binds to the surface and enters the
cytoplasm in small amounts but does not relocate to the nucleus. Trypan blue
marks dead cells.
F. localization of HAMLET in non-transformed human astrocytes. HAMLET enters the cytoplasm,
forming small aggregates, but does not relocate to the cytoplasm.
The cells remain viable for at least 24 h.

5. A B
Biopsy spheroids:
A. HAMLET B. α-lactalbumin

6.  Xenotransplantation
model
Human glioblastoma biopsy spheroids Infusion:HAMLET
α-lactalbumin

7. A
B
 Tumour Progression:
A. α-lactalbumin B. HAMLET

8.  Tumor volume and
survival
Tumor volume(mean) Pressure
symptoms(days)
α-lactalbumin 455 mm3 59
HAMLET 63 mm3 76
p< 0.01 p < 0.01

9.  Apoptosis induction in human
GBM biopsy spheroids.

10.  Therapeutic effect of HAMLET

13.  Thank you…CANCER RESEARCH by Walter Fischer, Lotta Gustafsson, Ann-Kristin
Mossberg, Janne Gronli, Sverre Mork, Rolf Bjerkvig, and
Catharina Svanborg1
Institute of Laboratory Medicine, Department of Microbiology, Immunology and
Glycobiology, University of Lund, Sweden; Department of Neurosurgery,
Haukeland University
Hospital, Bergen, Norway; Department of Physiology, University of Bergen,
Bergen, Norway; Department of Pathology, Haukeland University Hospital, Bergen,
Norway; and
NorLux Neuro-Oncology, Department of Biomedicine, University of Bergen and
Centre de Reserche Public de la Sante´ Luxembourg, Bergen, Norway