Differentiation Therapy has built a unique bridge between cell proliferation and differentiation for malignant cells which gave hope for researches in finding cure for cancers. These slides shows a summation of a pre-done video in explaining differentiation therapy and its clinical applications specifically on pediatric neuroblastoma and acute promyelocytic leukemia. All information in slides were obtained from external sources as in the citation and reference list. *Slides prepared by: Anna Mayr, Lalitha Ambighai, Cheah Toh Yang, Romel Mario Soyza, Gayathri Nanayakkara and Stephanie Veter* P.S Comments and questions in regards to researched topic would be very much welcomed and appreciated. Thank you. Have a nice day. :)

1. Lalitha | Anna | Stephanie | Gayathri
Romel | Toh Yang
╦╤──╤╤──╤╤──╤╤──╤╤
(BIO 3113)
GENES & TISUE CULTURE
TECHNOLOGY
Supervisor: Dr. Yap Wei Hsum

2. What is differentiation therapy?
To force malignant cells in undergoing terminal differentiation
>>> When cells are differentiated, less proliferation <<<
Malignant cells are undifferentiated
➔ No uniformity in shape and function
➔ Metastasis leads to poor prognosis
Current cancer therapies Differentiation Therapy
1. Highly toxic & nonspecific
2. Healthy cells affected
3. Increasing need to devise milder
treatments for older patients with
cancer
1. A less toxic & targeted approach as
it does not destroy cells but restrains
their growth
2. To reduce “tumour burden”
- Number of cancer cells -
- Size of tumor -
(Brown and Hughes 2012 ; Genetics Home Reference 2011 ; Leszczyniecka et al. 2001 ; MedicineNet.com 2015)

3. Differentiation
Agent Types
Differentiation
Agent Examples
Usage Current Status
Interferons (IFN) IFN Alpha 1970s : HL- 60 myeloid
leukemia cell line maturation
Not being used due to
occurrences of
haematological toxicity
Butyrates Butyric acid 1970s : induce gene expression
via histone hyperacetylation
Ongoing research to stabilize
the molecule
Possible candidate drugs for
MDS therapy.
Plant derived
alkaloids
Homoharringtonine
(HHT)
1970s : treat haemopoietic
diseases such as chronic
myeloid leukemia via
macrophage maturation
Not being used. Induction
death following neutropenic
infections occurring in 13/28
cases.
(Valeria and Pierluigi 2008)

4. • result of a translocation between chromosomes 15 and 17
• break in chromosome 15 disrupts the promyelocytic leukemia (PML) gene which encodes a
growth suppressing transcription factor
• break in chromosome 17 interrupts the retinoic acid receptor alpha (RARa) gene which regulates
myeloid differentiation
• translocation creates a PML/RARa fusion gene
• produces abnormal protein- causing,
arrest of maturation in myeloid cell
• reduces terminal cell differentiation
• increased proliferation of promyelocytes.
most fatal type of acute leukemia
(LinkedIn Corporation 2015 ; MedicineNet.com 2015 ; Shomilie 2014 ; Zhao et al. 2015)

5. APL
Treatment
Development
&
Mechanism of
Action
(Brown and Hughes 2012 ;
LinkedIn Corporation 2015 ;
Novak et al. 2009 ;
Nerdy Science Blog 2015 ;
Ozpolat 2008 ;
Valerie and Hugues 2013 ;
Zhou et al. 2007)

6. APL Treatment Challenges (cause – ATRA/ ATO)
HIGHLY THREATENING TO HIGHLY CURABLE ?
1. DIFFERENTIATION SYNDROME 2. PSEUDOTUMOR CEREBRI
Symptoms:
• Unexplained fever
• Acute renal failure
• Weight gain
Characterized by:
• Increased intracranial pressure
• Severe headache
• Nausea
• Vision disturbance
Onset- 7 days after initiation, LIFE THREATENING, High leukocyte count !
Can Overcome? DOSAGE VARIATION
(Kumar 2014 ; LinkedIn Corporation 2015)

7. •Common extracranial solid tumor of infancy.
•MYCN gene provides instructions for making a protein
that plays an important role in the formation of tissues
and organs during embryonic development
•MYCN is an oncogene that is overexpressed in
approximately one quarter of cases of neuroblastoma
via the amplification of the distal arm of chromosome 2
•Tend to have rapid tumor progression and poor prognosis
(Gunes and Akcakus 2002 ; Meduweb 2009)

8.  13-cis-retinoic acid (cis-RA) used in current differential treatments
 Differentiation therapy that involves inducing differentiation in malignant cells, eventually leading
to cell growth arrest and apoptosis.
 Neurite outgrowth used to screen a library of Micro-RNA mimics to identify those capable of
inducing differentiation in Neuroblastoma cells
 miR-506-3p is the most potent
New Differentiation Agents Identified for
Treatment of Pediatric Neuroblastoma

9. How Micro-RNA’s Function?
(Toh 2015)

10. Micro-RNA’s can target several genes!
miR-506-3p is more potent than 13-cis-retinoic acid (cis-RA)
This method has fewer toxic side effects compared to conventional
chemotherapy
(Zhao et al. 2014)

11.  Differentiation therapy is a treatment for cancer which has
been around for more than 40 years
 Differentiation therapy is considered a better cancer
therapy due to its less toxicity
 ATRA was the 1st differentiation inducer discovered and
was very effective for APL
 Current research focus : miRNA
Neuroblastoma | Acute Myeloid Leukemia
CONCLUSION
DT
GOOD
(Palma et al. 2015)

12. Boltz, K 2014, Differentiation therapy holds promise as a targeted therapy for pediatric neuroblastoma, viewed 8 September 2015,
http://www.oncologynurseadvisor.com/web-exclusives/differentiation-therapy-holds-promise-as-a-targeted-therapy-for-pediatric-
neuroblastoma/article/340516/
Brown, G and Hughes, P 2012, Retinoid Differentiation Therapy for Common Types of Acute Myeloid Leukemia, viewed 8th September
2015, http://www.hindawi.com/journals/lrt/2012/939021/
Genetics Home Reference 2011, Acute Promyelocytic Leukemia, viewed 8 September 2015, http://ghr.nlm.nih.gov/condition/acute-
promyelocytic-leukemia
Gunes, M and Akcakus, T 2002, Congenital Neuroblastoma with Cutaneous Metastases, viewed 10th September 2015,
http://indianpediatrics.net/mar2002/mar-308.htm
Kumar, S 2014, Acute promyelocytic leukemia NCCN LATEST 2014 Guidelines, viewed 16th September 2015,
http://www.slideshare.net/SandeepBhagat3/acute-promyelocytic-leukemia-nccn-latest-2014-guidelines
Leszczyniecka, M, Roberts, T, Dent, P, Grant S and Fisher, PB 2001, Differentiation therapy of human cancer: basic science and clinical
applications, viewed 10th September 2015, http://www.ncbi.nlm.nih.gov/pubmed/11578655
LinkedIn Corporation 2015, All-trans retinoic acid related complications in a patient with acute promyelocytic leukemia, viewed 16
September, http://www.slideshare.net/satanicala/alltrans-retinoic-acid-related-complications-in-a-patient-with-acute-promyelocytic-
leukemia
Reference List

13. MedicineNet.com 2015, Definition of Differentiation Therapy, viewed 7th September 2015,
http://www.medicinenet.com/script/main/art.asp?articlekey=19760
Meduweb 2009, Adrenal gland, neuroblastoma, microscopic picture- Endocrine Pathology Atlas, viewed 10th September 2015,
http://www.meduweb.com/threads/5995-Adrenal-gland-neuroblastoma-microscopic-picture-Endocrine-Pathology-Atlas
Nerdy Science Blog 2015, Chemotherapy, viewed 16th September 2015, http://science.kukuchew.com/2008/09/29/chemotherapy/
Nowak, D, Stewart, D and Koeffler, HP 2009, Differentiation therapy of leukemia: 3 decades of development, viewed 7th September
2015, http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2943835/
Ozpolat, B 2008, Acute promyelocytic leukemia and differentiation therapy: molecular mechanisms of differentiation, retinoic acid
resistance and novel treatments, viewed 9th September 2015,
https://www.researchgate.net/publication/26625718_Acute_promyelocytic_leukemia_and_differentiation_therapy_molecular_mecha
nisms_of_differentiation_retinoic_acid_resistance_and_novel_treatments
Palma, CA, Sheikha, DA, Teck, KL, Bryant, A, Thi, TV, Vivek, J, Ma, DF 2014,’MicroRNA-155 as an inducer of apoptosis and cell
differentiation in Acute Myeloid Leukaemia’, Molecular Cancer, vol.13, no.1, viewed 14 September 2015,
http://link.springer.com/article/10.1186%2F1476-4598-13-79

14. Shomilie, HA 2014, The acute-leukaemias, viewed 16th September, http://www.slideshare.net/Hatam-Shomilie/the-acuteleukaemias
Toh, GT 2015, (5.2) Epigenetics Mechanisms & Effects in Human (II), PowerPoint slides, Taylor's University, Subang Jaya, Malaysia.
Valeria, S and Pierluigi, RF 2008, DIFFERENTIATION THERAPY OF MYELODYSPLASTIC SYNDROMES: FACT OR FICTION?, viewed 11th
September 2015, http://onlinelibrary.wiley.com/doi/10.1046/j.1365-2141.1998.00881.x/full
Valerie, LB and Hugues, T 2013, Retinoic acid plus arsenic trioxide, the ultimate panacea for acute promyelocytic leukemia?, viewed 8Th
September 2015, http://www.bloodjournal.org/content/122/12/2008?sso-checked=true
Zhao, GB, Zhang, J, Wang, ZY, Chen, SJ and Chen, Z 2007, Treatment of acute promyelocytic leukaemia with all-trans retinoic acid and
arsenic trioxide: a paradigm of synergistic molecular targeting therapy, viewed 7th September 2015,
http://www.ncbi.nlm.nih.gov/pubmed/17317642
Zhao, Z, Ma, X, Hsiao, TH, Lin, G, Kosti, A, Yu, X, Uthra, S, Chen, Y, Tomlinson, GE, Pertsemlidis, A, Du, L 2014, A high-content
morphological screen identifies novel microRNAs that regulate neuroblastoma cell differentiation, viewed 8 September 2015,
http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path%5B0%5D=1703&path%5B1%
5D=2148

15. THANK YOU!
SAYONARA…