Radiobiology I

Direct and Indirect action

The biologic effects of radiation result principally from damage to DNA which is the critical target The atoms of the target itself may be ionized or excited, thus initiating the chain of events that leads to a biologic change called direct action The atoms may interact with other atoms or molecule in the cell to produce free radicals that are able to diffuse far enough to reach and damage the critical target.

Direct and Indirect action

Critical Target Theory Direct action -occurs when the radiation is absorbed by a molecule known to be critical to maintain life of the cell e.g. DNA.This may initiate a series of events that lead to changes that may be lethal to the cell This is the dominant process for high LET radiation

Indirect action
Indirect action --occurs when radiation interacts with other molecules in the cell,most importantly water.The products of these interactions may then go on to interact with the DNA This is the dominant process for low LET radiation About two thirds of the biologic damage by xrays is caused by indirect action

How does the ionising radiation damage the cells?

Indirect action: Electrons produce free radicals which break chemical bonds and produce chemical changes Direct Action: Photon ejects an electron which produce a biological damage on the DNA

Free radicals
A free radical is a molecule or atom,which is not combined to anything (ie.free) and carries an unpaired electron in its outer shell,i.e.its looking for something to interact with, or in purely scientific terms, it is in a state associated with a high degree of chemical reactivity.

Free Radical..
For simplicity let us consider what happens if radiation interacts with water molecule (80% of a cell is composed of water. If the water molecule is ionised H2O =H2 O++e- ; H2 O is the water molecule H2O+ is an ion radical.

Free Radical..
Ion meaning it is electrically charged, because it has lost an electron and a radical because it has an unpaired electron in the outer shell,making it very reactive. Ion radicals have a short life,usually no more than 10-10 seconds, before they decay to form free radicals

 Free radicals are not charged, but do have an unpaired electron in the outer shell. The water ion radical can for example do the following:
H2 O+ +H2O =H3O+ +OH* H2 O+, H3O+ are the ion radicals OH*is a highly reactive hydroxyl radical, with 9 electrons, therefore one is unpaired. Hydroxyl radicals (OH*),are highly reactive and can go on to react with DNA.It is estimated that 2/3 of the x-ray to react with DNA.

Indirect action The Process

Incident X-ray photon Fast electron (e-) Ion Radical Free Radical Chemical changes from the breakage of bonds Biologic effects

In summary the Indirect action is as follows

Incident x-ray photon Fast electron (e-)-occurs in 10-15 seconds Ion radical -live about 10-10 seconds Free radical -live about 10-5 seconds Chemical changes from the breakage of bonds Biological effect -may be expressed in hours, days, months, years or not at all, depending on the consequences of the bonds broken.

The time scale

The Physics of the absorption process is over in 10-15 second; The chemistry takes longer because the lifetime of the DNA radicals is about 10-3 to 10-5 second; The biology takes days to months for cell killing, years for carcinogensis, and generations for heritable damage.

Single & Double strand break

When cells are irradiated with x-rays, many breaks of single strand occur. These single-strand breaks are of little biologic consequence as far as cell killing is concerned as they are repaired readily using the opposite strand as a template

Double strand break

If both the strands of the DNA are broken and the breaks are well separated, repair again occurs readily, because the two breaks are handled separately. If the breaks in the two strands are opposite one another, or separated by only a few base pairs this may lead to a double-strand break. That is the piece of chromatin snaps into two pieces

Single & Double strand break

 A double strand break is believed to be the most important lesion produced in chromosomes by radiation. The yield in irradiated cells is about 0.04 times that of single strand breaks. The double strand break is induced linearly with dose

What is cell death?

For differentiated cells that do not proliferate, such as nerve, muscle, or secretory cells, death can be defined as loss of specific function. For proliferating cell such as stem cell in hematopoietic system or the intestinal epithelium, loss of capacity for sustained proliferation loss of reproductive integrity

THESE DAMAGES CAN LEAD TO

Slowdown in the cell synthesizing copies of its DNA, so that there is a delay in one cell dividing into two cells Delays (to allow repair) as the cell progresses towards its next cell division (delay in cell cycle progression) Decrease in the overall rate of cell proliferation (increase in cell number) of a population of cells Death of the cell Mutation of the cell Changes in the cell which will make it cancer-like (called cell transformation)

THE TYPES OF DAMAGE TO THE DNA INCLUDE

DNA Single Strand Breaks DNA Double Strand Breaks Sugar Damage Base Damage Local Denaturation (Separation of the 2 strands) DNA-DNA Cross-links DNA-Protein Cross-links

DIFFERENT TYPES OF CELL DEATH

General Description INTREPHASE DEATH: Death before the next cell division, or death of a cell that does not divide REPRODUCTIVE DEATH: Death of the cell (and its daughter cells) after one or more cell divisions Specific Description NECROSIS: Death of a contiguous (touching) field of cells Does not require energy; the contents of the cells leak into the surrounding tissue and blood supply CELL LYSIS: The cell simply bursts open, releasing its contents APOPTOSIS (or Programmed Cell Death): This type of death is under genetic control (specific genes must be present and active or inactive). It requires energy, and when the cells die, DNA fragments of specific sizes, and the contents of the cells, are encapsulated in membranes as small vesicles.

CHROMOSOME ABERRATIONS
Types, Dose and Dose Rate Dependence Ionizing radiation exposure results in many different types of aberrations, with the type depending on where the cell is in relation to its next division (position in its cell cycle). The most commonly measured types of aberrations are ring and dicentric aberrations, which can be used for biological dosimetry after an acute whole-body exposure above 10 25cGy (within a defined period after the exposure) There are many other types of aberrations that can occur, and if they (like the ring and dicentric aberrations) are obvious upon microscopic observation, the cell with those aberrations would likely have died. Certain kinds of chromosome aberrations, as well as genetic mutations of the DNA in the chromosomes, can be associated with causing cancer.

THE RELATIVE RADIOSE SITIVITY OF THE CELLS I THE BODY

Fully differentiated, functional and non-dividing cells (e.g, nerve cells, muscle cells) are RADIORESISTANT Partially differentiated cells that can be called upon to divide again (e.g., liver cells, glandular cells) are somewhat less radioresistant Cells which can divide but lend support to the other cells in a tissue (e.g. endothelial cells lining the blood vessels, fibroblasts of the connective tissue) are intermediate in radiosensitivity

RADIATION AND CANCER

Cancer resulting from exposure of cells to ionizing radiation is a stochastic or probability phenomena The outcome is either yes or no, and there is no threshold of dose below which ionizing radiation cannot induce cancer The types of cancers due to exposure of a large number of persons to ionizing radiation include both blood cancers and solid tumors The relationship may be either linear or linear-quadratic, depending on the type of cancer (e.g., for blood cancers, the incidence increases in a linear quadratic manner with dose, while for solid tumors, the increase is linear with dose, and fractionation does not decrease the risk

Cell survival curve

The cell survival curve is plotted on a semi log plot For low doses for sparsely ionizing radiations, the survival curve starts out straight on the semi log plot with a finite initial slope i.e. the surviving fraction is an exponential function of dose

Linear quadratic theory

Cell is inactivated by DSB Single hit, single lesion [random process], governed by Poisson statistics S = exp (-D)
S - surviving fraction of cells - average probability per unit dose that this will occur D - dose delivered

Linear quadratic theory

2 separate ionising events, probability of one interaction causing one lesionis linearly proportional to dose, as is mean probability of second particle doing the same Mean probability of both events is D2 - mean probability per unit dose squared that such complementary events will occur

Linear quadratic theory

In general cell survival is described by:
S = exp (-D -D2)

damage (irrepairable) damage (repairable)

NB of special interest is when D = -D2 i.e. the curviness of cell survival curve

/ ratio
High / [straighter curve], characteristic of cell with little repair capability e.g. tumour cells [from 5 - 20 Gy] Low / [more curved], characteristic of high repair potential e.g.late responding normal tissue [1-4 Gy] This difference in cell surv ival curves provides rationale for fractionated radiation therapy treatment and explains radiobiological advantage

MAMMALIAN CELL SURVIVAL CURVE

Survival vs. Dose Shoulder Region Shows accumulation of SUBLETHAL DAMAGE The larger the shoulder region, the more dose will initially be needed to kill the same proportion of cells Beyond the Shoulder Region The Do Dose, or the inverse of the slope of the curve, indicates the relative radiosensitivity. The smaller the Do dose, the greater the radiosensitivity

Cell Survival
S=e-D+D2 D=D2 D= /

MAMMALIAN CELL SURVIVAL CURVE

The Effect of Lowering the Dose Rate

Cell cycle and Radiosensitivity

Cells are most sensitive at or close to mitosis Resistance is usually greatest in the latter part of S phase If G1 phase has an appreciable length a resistant period is evident early in G1, followed by a sensitive period toward the end of G1 G2 phase is usually sensitive, perhaps as sensitive as M phase

The stages of the mitotic Cycle

M

G2

G1

S (DNS Synthetic phase)

Cell survival curve for various Phases of Cell cycle