Study of effect of HDAC inhibitor on regeneration in Eisenia fetida

Study of effect of HDAC inhibitor on regeneration in Eisenia fetida
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We maintained cup cultures for earthworms by using cellulose as their source of food. Our objective is to check the effect Valproic acid which is an HDAC inhibitor on the regeneration in earthworms.
We made 3 groups each containing 5 earthworms. Group A contains the control in which the earthworms are amputated at 40th segment and both the parts are kept seperately in different cups and is named as CE1A & CE1P meaning control earthworm 1 anterior part after amputation.
Similarly group B contains the worms amputated at 40th segment but treated with 1mM concentration of VPA at the site of amputation and named EE1A(1mM) & EE1P(1mM) and so on.
Similarly group C contains the worms amputated at 40th segment but treated with 2mM concentration of VPA at the site of amputation and named EE1A(2mM) & EE1P(2mM) and so on

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@bhakta_anupriya why are you using Valproic acid?
what is your expectation in control untreated vs treated ones?
what do you mean by HDAC inhibitor?
what is the research question you are addressing by using VPA?

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@jaikishan Valproic acid is an HDAC inhibitor.
My expectation in the control ones are they will regenerate faster as compared to the treated ones.
HDAC inhibitors are those compounds which inhibit histone deacetylation by inducing cell cycle arrest.
By using VPA we are trying to study the effect of VPA on cell division which will be later used for cancer treatments.

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Why were those particular concentrations of Valproic acid used?

@Akshitha The identification of VPA as an inducer of
both cell differentiation (20) and apoptosis argues that this
drug might play a dual role in thyroid cancer management;
VPA increases the efficacy of radiometabolic therapy, pro-
moting iodine reuptake by tumor cells, and affects tumor
growth by acting on the cell cycle and on cell death.
VPA levels reached in patients treated for epilepsy are
usually not above 100 g/ml (0.7 mm). Only limited toxicity
occurs when the concentration is below 3.1 mm, and severe
side effects develop when the concentration is above 5.9 mm;
1 mm VPA, which is the expected plasma level for use in
treating poorly differentiated thyroid cancer, is just above
therapeutic levels for epilepsy and thus appears clinically
achievable. Translational and clinical studies will ultimately
determine the clinical utility and safety of VPA as an option
for the treatment of poorly differentiated thyroid cancers,
which are known not to respond to conventional therapy

They have mentioned that 1mM is the maximum limit for humans and they have got the results that 3mM is the maximum conc where cell will go under apoptosis and will not further be divided.
So we are considering 3mM as a control where we will observe the inhibition of regeneration process where as 1mM might be and might not inhibit the regeneration because it’s maximum limit is for humans and we are dealing with earthworms.

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Interesting :smile:
Waiting for updates.

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So, how are you planning to prepare the 1mM/3mM VPA solutions?

Why will VPA treated earthworm will not regenerate?
What is your explanation @bhakta_anupriya?

Do you have any reference from literature on already known role of VPA in regeneration? please provide reference with link

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They will regenerate but the rate will be slower as compared to the untreated ones. @jaikishan

What do you mean by slower?
Why slower? What will VPA actually do ?
how much time? What is your expectation? @bhakta_anupriya

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Kindly check the link below!