Erythropoietic protoporphyria

IDENTIFYING THE PROBLEM

Problem to be solved
What are porphyrias?
Porphyrias are rare disorders that mainly affect the skin or nervous system. These disorders
are usually inherited, meaning they are caused by gene mutations passed from parents to
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children.

If you have porphyria, cells fail to change chemicals in your body—called porphyrins and
porphyrin precursors—into heme, the substance that gives blood its red color. When these
chemicals build up in your body, they cause illness. Depending on the type of porphyria you
have, porphyrins or porphyrin precursors may build up in the liver or the bone marrow. Bone
marrow is the spongy tissue inside most of your bones.

What are the types of porphyria?

Experts often divide porphyrias into two groups—acute porphyrias and cutaneous
porphyrias—based on whether they primarily affect the nervous system or the skin.

Acute porphyrias

Four types of acute porphyrias affect the nervous system. Two of those types can also affect
the skin. Symptoms for acute porphyrias develop over hours or days and last for days or
weeks.

Table 1. Types of acute porphyria

Where Porphyrins or
Parts of the Body
Type of Acute Porphyria Porphyrin Precursors
Affected
Build Up

acute intermittent porphyria nervous system liver

nervous system and

variegate porphyria liver
skin

nervous system and

hereditary coproporphyria liver
skin

delta-aminolevulinic acid (ALA) dehydratase

nervous system liver
deficiency porphyria

Cutaneous porphyrias

Four types of cutaneous porphyrias affect only the skin and cause chronic, or long lasting,
symptoms. People with cutaneous porphyria may develop skin symptoms—such as blistering
or pain—after their skin is exposed to sunlight.
Table 2. Types of cuteaneous porphyria

Parts of the Body Where Porphyrins

Type of Cutaneous Porphyria
Affected Build Up

porphyria cutanea tarda skin liver

protoporphyrias: erythropoietic
skin bone marrow
protoporphyria and x-linked protoporphyria

congenital erythropoietic porphyria skin bone marrow

hepatoerythropoietic porphyria skin liver

How common are porphyrias?

Porphyrias are rare diseases. Studies suggest that all types of porphyrias combined affect
fewer than 200,000 people in the United States.1

The most common type of acute porphyria is acute intermittent porphyria.

The most common type of cutaneous porphyria—and the most common type of porphyria
overall—is porphyria cutanea tarda, which affects about 5 to 10 out of every 100,000 people. 2

The most common type of porphyria in children is a cutaneous porphyria called

erythropoietic protoporphyria.3

Who is more likely to get porphyria?

Acute porphyria is more common in females than in males and often begins when people are
between the ages of 15 and 45.4

Among types of cutaneous porphyria, porphyria cutanea tarda most often develops in people
older than age 40, usually men.2 For other types of cutaneous porphyria, symptoms often
appear in early childhood.

What are the complications of porphyrias?

Different types of porphyrias may lead to different complications.

Liver problems

Several types of porphyrias can cause liver problems. Acute porphyria increases the chance of
developing liver cancer . Porphyria cutanea tarda can damage the liver and increase the
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chance of developing cirrhosis and liver cancer. Some people with protoporphyria also
develop liver damage and cirrhosis, and up to 5 percent of people with protoporphyria
develop liver failure.5

In people with protoporphyria, bile carries extra porphyrins from the liver to the gallbladder,
which may lead to gallstones that are made of porphyrins.
Anemia

Two types of cutaneous porphyria, congenital erythropoietic porphyria and, less commonly,
hepatoerythropoietic porphyria, may cause severe anemia. These diseases may also cause
the spleen to become enlarged, which can make anemia worse.

High blood pressure and kidney problems

People with acute porphyria have an increased chance of developing high blood
pressure and chronic kidney disease, which can lead to kidney failure.

Porphyrins and porphyrias Porphyrias are a family of inherited diseases, each associated
with a partial defect in one of the enzymes of the heme biosynthetic pathway (1) (Figure 1).
As a consequence, each porphyrinopathy shows specific metabolic abnormalities reflected
by diagnostic laboratory findings in blood, urine and feces. Based on the main clinical
manifestation, porphyrias are currently classified into acute and non-acute(2) (Table 1).
In six of the eight described porphyrias, the main clinical feature is skin photosensitivity

brought about by the action of light on porphyrins, which are deposited in the upper
epidermal layer of the skin. Porphyrins absorb radiant energy intensively in the UV region, at
their Soret band (400-410 nm) and to a lesser extent in the long visible bands (580-650 nm),
resulting in transitions to excited electronic states (Figure 2). In the cutaneous porphyrias,
skin lesions are due to these properties of porphyrins. The excited porphyrin may react
directly with biological structures (type I reactions) or with molecular oxygen, generating
excited singlet oxygen (type II reactions).
Singlet oxygen in turn damages tissues through several mechanisms, including oxidation of
membrane lipids and amino acids in proteins, cross-linking of proteins and oxidative damage
to nucleic acids with the consequent disturbance of the normal functioning of cellular
activities or organelles. In addition to singlet oxygen, the other reactive oxygen species
(ROS), O2 ·- , H2O2, and · OH are also formed, which are also important in the progress of
porphyrin photosensitization

Further studies showed that the key determinants for the photodynamic effectiveness lie in
the solubility properties of the porphyrin sensitizer relative to the hydrophilic or
hydrophobic nature of the target biomolecules, with PROTO being most phototoxic to
lipophilic structures, such as erythrocyte membranes, mitochondrial preparations and
membrane

X-linked dominant protoporphyria (XLDPP) protoporphyrin ring

SELECTION OF SOLUTION (LAST DANCE)

Another promising avenue for regulating hormonal balance with nano-devices is through the
development of nanoscale drug delivery systems. These systems can be designed to target
specific hormone-producing cells or glands, delivering therapeutic agents with high precision
and minimizing off-target effects. By incorporating energy-responsive elements into these
nanoscale delivery systems, researchers aim to achieve on-demand control over hormone
release and activity.
Combination of absorbent , Nanoscale Drug Delivery System, and nano sensor by addition to
the compatible coating.

ABSORPTION OF )ROS( BY NANOSCALE PARTICLES.

The absorption of reactive oxygen species (ROS) by nanoparticles is a topic of significant

interest due to its potential applications in various fields such as medicine, environmental
remediation, and catalysis. Reactive oxygen species are highly reactive molecules containing
oxygen, such as superoxide anion (O2•−), hydroxyl radical (•OH), and hydrogen peroxide
(H2O2). These species play crucial roles in biological processes and are also implicated in
oxidative stress-related diseases.

Nanoparticles have been investigated for their ability to scavenge or absorb ROS due to
their unique physicochemical properties, including high surface area, tunable surface
chemistry, and size-dependent reactivity. Furthermore, the size and morphology of
nanoparticles also influence their ability to absorb ROS. Smaller nanoparticles generally
exhibit higher surface-to-volume ratios, which can enhance their reactivity towards ROS.
Additionally, the shape of nanoparticles can impact their ROS absorption capacity. For
example, nanomaterials with high aspect ratios or specific crystal facets may exhibit
enhanced ROS scavenging properties due to their unique surface structures.

In addition to direct interactions with ROS, nanoparticles can also possess intrinsic
antioxidant properties that contribute to ROS absorption. Certain nanomaterials, such as
carbon-based nanomaterials (e.g., graphene oxide, carbon nanotubes) and polymeric
nanoparticles, contain functional groups or chemical moieties that can act as antioxidants by
donating electrons or hydrogen atoms to ROS, thereby mitigating their damaging effects.
Moreover, some nanoparticles can encapsulate or immobilize antioxidant compounds within
their structures, providing sustained ROS-scavenging capabilities. It is important to note that
the absorption of ROS by nanoparticles is a complex process influenced by various factors
including nanoparticle composition, size, surface chemistry, and environmental conditions.

Mechanism
Chosen method :
Internalization: ROS can be internalized within the particles, where they can be trapped
and neutralized by the particle’s matrix. This mechanism is more effective for particles
with high internal volumes, such as liposomes and nanoparticles made of polymers.

When ROS are generated in a system, such as in cells or in the environment, they can
interact with various particles present in that system. These particles may include
biological components such as cell membranes or organelles, as well as environmental
particles like soil minerals or atmospheric aerosols. When ROS come into contact with
these particles, they can undergo internalization, becoming trapped within the particle’s
structure.

The internalization of ROS within particles serves several important functions. Firstly, it
can help to mitigate the damaging effects of ROS. ROS are highly reactive molecules that
can cause oxidative damage to biological structures and environmental components. By
 being internalized within particles, ROS are effectively sequestered and neutralized,
preventing them from causing harm.

The process of internalization is influenced by various factors including the properties of

the particles involved (such as surface chemistry and porosity), the specific types of ROS
present, and the environmental conditions (e.g., pH, temperature). Understanding the
mechanisms and implications of internalization is important for fields such as
environmental science, toxicology, and nanotechnology.

Factor effecting absorption

Several factors can affect the absorption of ROS by nano-scale particles, including:

1. Particle size: The size of the particles can affect their ability to absorb ROS. Smaller
particles have a larger surface area-to-volume ratio, making them more effective at
absorbing ROS.
2. Particle material: The material composition of the particles can affect their ability to
absorb ROS. Particles made of materials with high surface areas, such as carbon
nanotubes and graphene, are more effective at absorbing ROS.
3. Concentration of ROS: The concentration of ROS in the environment can affect the
ability of nano-scale particles to absorb ROS. Higher concentrations of ROS can lead
to more efficient absorption.
4. Temperature: The temperature of the environment can affect the absorption of ROS
by nano-scale particles. Higher temperatures can increase the rate of ROS
absorption.

BIOLOGICAL PROCESSES BY ROS

Once trapped within a particle, ROS may undergo chemical reactions or transformations
that alter their reactivity or toxicity. For example, certain minerals in soil have been shown
to catalyze the conversion of superoxide radicals into less reactive species.
Enzymes are biological molecules that catalyze specific biochemical reactions, playing a
crucial role in maintaining the homeostasis of various physiological processes. One of the
essential aspects of enzyme function is the balance between the activity of different
enzymes, which is critical for maintaining cellular homeostasis. This balance is often
disrupted in various diseases, and understanding the mechanisms that regulate enzyme
activity is essential for developing therapeutic strategies.

ROS BOLD (Reactive Oxygen Species-Based Optimization of Enzyme Localization and

Dynamics) is a novel approach that aims to enhance enzyme balance by modulating the
activity of enzymes using reactive oxygen species (ROS). ROS are highly reactive molecules
that can damage cellular components, but they also play a crucial role in cellular signaling
and homeostasis.
How ROS BOLD Works
ROS BOLD works by using ROS to fine-tune the activity of enzymes, thereby maintaining the
balance of enzymatic activity. This approach involves the following steps:

1. Identification of key enzymes: The first step is to identify the key enzymes that are
involved in the desired physiological process. These enzymes are often critical for
maintaining cellular homeostasis, and their dysregulation can lead to various
diseases.

2. Design of ROS-sensitive enzyme variants: Once the key enzymes have been
identified, the next step is to design ROS-sensitive variants of these enzymes. These
variants are engineered to be more sensitive to ROS, which allows for more precise
regulation of their activity.

3. ROS treatment: The ROS-sensitive enzyme variants are then treated with ROS,
which fine-tunes their activity. The amount of ROS used is critical, as excessive ROS
can damage cellular components, while too little ROS may not have any effect.

4. Monitoring of enzyme activity: The activity of the enzymes is monitored using

various biochemical assays to ensure that the desired level of activity is achieved.

Benefits of ROS BOLD

The ROS BOLD approach has several benefits, including:
1. Precise regulation of enzyme activity: ROS BOLD allows for precise regulation of
enzyme activity, which is critical for maintaining cellular homeostasis.
2. Minimized side effects: The use of ROS-sensitive enzyme variants minimizes the risk
of side effects, as the activity of the enzymes can be fine-tuned to achieve the
desired outcome.

3. Flexibility: ROS BOLD can be used to regulate the activity of multiple enzymes
simultaneously, allowing for a more comprehensive approach to maintaining cellular
homeostasis.

This step will be completed with nanoscale drug delivery systems

nanoscale drug delivery system

SAFETY OF USING NANO-TECH INSIDE HUMAN BODY

Impacts

The impacts of nano-scale particles on the human body can be diverse and are influenced
by various factors such as size, shape, surface chemistry, and composition. When these
particles are inhaled, ingested, or come into contact with the skin, they can interact with
biological systems in different ways.
 1. Inhalation: Nano-scale particles can enter the body through inhalation and reach
the respiratory system. Due to their small size, they can penetrate deep into the
lungs and potentially cross into the bloodstream. This raises concerns about
respiratory diseases, inflammation, and potential long-term effects on lung function.

2. Ingestion: When nano-scale particles are ingested, they may interact with the
gastrointestinal tract and be absorbed into the bloodstream. This raises concerns
about their potential systemic distribution and accumulation in various organs.
3. Skin Contact: Nano-scale particles can also penetrate the skin barrier and enter the
body through dermal exposure. This raises concerns about skin irritation,
sensitization, and potential systemic effects.

Once inside the body, nano-scale particles can interact with cells, tissues, and organs,
potentially leading to oxidative stress, inflammation, genotoxicity, and other adverse
effects. Furthermore, their small size allows them to reach locations that larger particles
cannot access. This unique ability raises concerns about their persistence and potential
long-term effects within the body.

Research has shown that some nano-scale particles can persist in the body for extended
periods. Their ability to evade clearance mechanisms and accumulate in certain tissues
raises concerns about chronic exposure and potential health risks over time.
The duration of nano-scale particle persistence in the body depends on various factors
such as their chemical composition, surface modifications, and interactions with biological
systems. Some studies have indicated that certain types of nano-scale particles can remain
in the body for prolonged periods, potentially leading to bioaccumulation and chronic
toxicity.

It’s important to note that not all nano-scale particles exhibit the same behaviour inside the
body. Some are designed to biodegrade or be cleared from the body through natural
processes. However, understanding the fate of these particles inside the body is essential for
assessing their long-term safety.

Safety precautions
The parameters that have been considered during the design of our solution was as follows

1. Material Selection: The choice of materials for nanoparticles is crucial in determining

their safety and longevity. Biocompatible materials that are non-toxic to the human body
are preferred for designing safe nanoparticles. For example, biodegradable polymers such as
poly(lactic-co-glycolic acid) (PLGA) and chitosan have been widely used in nanoparticle
formulations due to their low toxicity and ability to degrade into non-toxic by-products. In
addition, inorganic materials such as gold and silica have also been explored for their
biocompatibility and stability. By selecting appropriate materials, the risk of adverse effects
on the human body can be minimized, while ensuring the longevity of the nanoparticles.
2. Surface Modification: Surface modification plays a critical role in enhancing the stability
and biocompatibility of nanoparticles. Functionalizing the surface of nanoparticles with
biocompatible coatings or ligands can improve their stability in physiological environments
and reduce potential interactions with biological components that may lead to adverse
effects. For instance, polyethylene glycol (PEG) coating has been widely used to impart
stealth properties to nanoparticles, reducing their recognition by the immune system and
prolonging their circulation time in the body. Surface modification can also help prevent
aggregation of nanoparticles, which is essential for maintaining their stability over an
extended period.
3. Controlled Release Mechanisms: In drug delivery applications, designing nanoparticles
with controlled release mechanisms can contribute to their long-term effectiveness while
minimizing potential toxicity. Encapsulation of therapeutic agents within nanoparticles
allows for sustained release over time, reducing the need for frequent dosing and
minimizing potential side effects associated with high drug concentrations. Moreover,
stimuli-responsive nanoparticles can be engineered to release their cargo in response to
specific triggers such as pH, temperature, or enzymatic activity, providing precise control
over drug release while ensuring safety and efficacy.
4. Size and Shape Optimization: The size and shape of nanoparticles can significantly
influence their behavior in biological systems. Studies have shown that smaller nanoparticles
may exhibit prolonged circulation times in the body due to reduced clearance by the
reticuloendothelial system, while certain shapes such as spheres or rods may offer
advantages in terms of cellular uptake and biodistribution. By optimizing the size and shape
parameters, it is possible to design nanoparticles that not only last longer in the body but
also minimize potential adverse effects on human tissues.
5. Biodegradability and Clearance: Incorporating biodegradable components into
nanoparticle formulations can facilitate their gradual degradation into non-toxic byproducts,
allowing for safe clearance from the body over time. This is particularly important for long-
term applications where persistent accumulation of nanoparticles could pose risks.
Strategies such as designing hybrid nanoparticles with both biodegradable and non-
biodegradable components or engineering stimuli-responsive degradation mechanisms can
enable controlled breakdown of nanoparticles without eliciting harmful effects on the
human body.