Gene & Cell Therapy

antonia.follenzi@med.uniupo.itVia Solaroli,17 – NovaraHealth Sciences Dept.

Tel. 0321-660674

REGENERATIVE MEDICINE

Definition of stem cells: stem cells are unspecialized cells that have two defining properties: the ability to differentiate into other cells and the ability to self-regenerate/self-renew

This cellCan form the

Embryo and placenta

This cellCan just form the

embryo

Fully mature

Stem Cell Types Based on The Ability to Differentiate

• Totipotent all cell types. E.g.: zygote• Pluripotent all three germ layers. E.g.:

human embryonic stem cells• Multipotent many cell types. E.g.:

hematopoietic stem cells• Unipotent can produce only one cell

type, but have the property of self-renewal which distinguishes them from non-stem cells

POTENCYDEVELOPMENT

Cloning

Regeneration

Inducedreprogramming

The progressive loss of potency

Differentiation of Human Tissue

Simposium Stem Cell di FKUI, ULTAH Kalbe 40 tahun, 2 Sept.2006

Inner cell mass

Day 5 after conception

Adult Stem Cells: Bone Marrow

Plasticity of Adult Stem Cells

Oct3/4Sox2Nanog

Ecat1Ecat8Dppa2Dppa3Dppa4Dppa5Dnmt3lFbx15Gdf3Sox15Fthl17Sall4Rex1Utf1Grb2Tcl1

-cateninKlf4c-MycStat3E-Ras

Induced reprogramming of mouse cells

Takahashi & Yamanaka, Cell, 2006

Induced reprogramming of mouse cells

Induced reprogramming of human cells

Modeling human disease with iPS cells

Modeling human disease with iPS cells

Modeling human disease with iPS cells

Modeling human disease with iPS cells

Modeling human disease with iPS cells

Treating human disease with iPS cells

Self-renewal> 20 passagesKaryotypic stability

PluripotencyExpression of pluripotency-associated markersIn vitro differentiationTeratoma formation

MolecularDNA fingerprintingIntegration of reprogramming transgenesSilencing of reprogramming transgenesReprogramming of gene expression profileReprogramming of DNA methylation profile

Self-renewal> 20 passagesKaryotypic stability

PluripotencyExpression of pluripotency-associated markersIn vitro differentiationTeratoma formation

MolecularDNA fingerprintingIntegration of reprogramming transgenesSilencing of reprogramming transgenesReprogramming of gene expression profileReprogramming of DNA methylation profile

Criteria for defining bona fide iPS cells

Simposium Stem Cell di FKUI, ULTAH Kalbe 40 tahun, 2 Sept.2006

Research & Clinical Applications of Cultured Stem Cells

• Functional genomic studies gene therapy

• Study of biological processes development of the organism & progress of cancer

• Drug discovery & development to see the properties of the drugs to differentiated cells

• Cell-based regenerative therapy

Diseases will potentially be treated

• Parkinson's and Alzheimer's diseases

• Spinal cord injury • Stroke • Burns • Heart disease • Diabetes

• Muscular dystrophy• Osteoporosis injuries• Cirrhosis hepatis• Leukemia• Sickle cell anemia• Osteoarthritis • Rheumatoid arthritis • Cancer

Stem Cell Characteristics Make Them Good Candidates for Cell-based Therapies

• Potential to be harvested from patients• High capacity of cell proliferation in culture to

obtain large number of cells from a limited source

• Ease of manipulation to replace existing non functional genes via gene transfer methods

• Ability to migrate to host's target tissues, e.g. the brain

• Ability to integrate into host tissue and interact with surrounding tissue

Paracelsus

• The fundamental theory behind organ extract and The fundamental theory behind organ extract and cell therapy is the principle cell therapy is the principle

‘‘Similia Similibus curanturSimilia Similibus curantur’ or ’ or ‘Like Cures Like’‘Like Cures Like’, ,

as stated by Paracelsus, a Swiss physician and as stated by Paracelsus, a Swiss physician and philosopher of the 16philosopher of the 16thth century. Paracelsus and century. Paracelsus and many other early physicians believed that the best many other early physicians believed that the best way to rebuild or revitalise ill organs or ageing way to rebuild or revitalise ill organs or ageing tissue was to use healthy living cells of the same tissue was to use healthy living cells of the same tissue type. tissue type.

In 1492, Pope Innocent VIII is said to have received, at the behest of a Jewish physician, a transfusion of the blood of three ten year old boys, each of whom was paid a ducat andall of whom died. Probably the blood was drawn, but wasintended to be taken orally. Indeed, there is no reliable evidence that the sickly pope accepted the blood at all.

This story has been told and retold over the last half millennium. It is most likely apocryphal and has the flavor of an early urban legend in its details and its anti-Semitic and anti-Catholic overtones.

“First Transfusion” Myth

Richard Lower (1631-1691)

Richard Lower is credited with performing, in 1665, the first authentic blood transfusion (animal to animal).

He kept exsanguinated dogs alive by connecting the carotidartery of the donor dog to the jugular vein of the recipient dog.

Blood Transfusion

• 1818 - James Blundell, a British obstetrician, performed the first successful transfusion of human Blood to a patient for the treatment of postpartum hemorrhage.

– Using the patient's husband as a donor, he extracted a small amount of Blood from the husband's arm and, using a syringe, he successfully transfused the wife. Between 1825 and 1830, he performed ten documented transfusions, five of which proved beneficial to his patients, and published these results.

Karl Landsteiner1930 Nobel Prize Laureate In 1900, Landsteiner

showed that serum fromsome individuals couldagglutinate or hemolyzethe red blood cells of certain, but not all, other individuals. The serum ofthe latter would likewiseagglutinate the red bloodcells of the former. Still other individuals’ red cells were unaffected by the serum from either of these.He named these threedifferent types A, B, andC. Today these are typesA, B, and O.

Dr. Alexis Carrel

• In the late 19th century, the French Nobel laureate discovered the potentially immortal nature of cells by keeping alive fragments of a chicken heart 25 years after the fowl had died.

E. Donnall Thomasthe father of bone marrow transplantation

The spirit of a pioneer

• 1956 – The First Transplantation Between Identical Twins was performed in Cooperstown, New York– Thomas reported that total body irradiation followed

by infusion of marrow from an identical twin could result in complete remission of leukemia. 

1968 – First Bone Marrow Transplant Between Siblings

In 1968, however, enough was known about the HLA system for a transplant between siblings to occur.  Dr. Robert A. Good performed the first successful transplant of this sort at the University of Minnesota: 

- the recipient was a four-month-old boy who had inherited severe combined immunodeficiency syndrome (“bubble boy syndrome”) and the donor was his eight-year-old HLA-matched sister.  The disease had previously killed 11 male children in the boy’s family. 

1973 – First Unrelated Bone Marrow Transplant

• Expanding knowledge of the HLA-system allowed a team at Memorial Sloan-Kettering Cancer Center in New York City to perform the first unrelated bone marrow transplant in 1973 on a five-year-old patient suffering from sever combined immunodeficiency syndrome.  The matched donor was found in Denmark through the Blood Bank at Rigshospitalet in Copenhagen.  The patient received multiple infusions of marrow, and after the seventh transplant, engraftment was achieved and hematologic function became normal.

Dr. Thomas, wins the 1990 Nobel Prize in Physiology or Medicine for his pioneering work in the area of organ

and cell transplantation. 

October 2012 Dr Thomas died