jl club

1. By Luke Lomas
22/1/2019
JOURNAL CLUB:
UVEAL
MELANOMA

2. JOURNAL ARTICLE: NESTIN EXPRESSION IN
PRIMARY AND METASTATIC UVEAL
MELANOMA
–
POSSIBLE BIOMARKER FOR HIGH-RISK UVEAL
MELANOMA
 Djirackor, L., Shakir, D., Kalirai, H., Petrovksi, G. and Coupland, S. (2017).
Nestin expression in primary and metastatic uveal melanoma. Acta
Ophthalmologica, 95.

3. ABOUT THE PAPER
 In this paper they examine the putative stem cell
marker nestin, comparing the expression of nestin in in
primary uveal melanoma (PUM) and metastatic uveal
melanoma (MUM).
 Nestin expression is associated to the latter stages of
tumour progression where poor survival rate is
predicted.
 One problem about the paper is that it states that
there is very little information relating uveal melanoma
to nestin expression.
 CD133 was used as a marker as it is nestin positive. In
the blood of patients with MUM circulating tumour
cells which correlated to the nestin positive tumour
cells that were in the tissue (CD133).
 In the study immunohistochemistry was used in
correlation with morphology, genetic features and the
patients eventual outcome. (H&E) Staining.
 6 uveal melanoma (UM) cell lines used.

4. METHODOLOGY
In this study the methodology used was immunohistochemistry and that
alone. The samples were stained with mouse antinestin and then incubated
with horseradish peroxidase to detect the bound antibody and fluoresce,
Tissue sections were stained with haematoxylin. Normal colon sections
were used as a positive control to validate the IHC run and then images
were taken.
One positive of this study was that there was a large number of samples of
both PUM (141 patients) a 26 (Hepatic metastases MUM). Which supports
this study.
10% Nestin was used as a threshold if above this considered a valid
indicator.

5. From the results it is clear that when
there is greater than 10% Nestin the
basal diameter, ciliary body
involvement and extraocular
extension is greater. The percentage
of necrosis is also greater. When the
nestin levels are greater the
chromosome changes are what you
would expect to see in typical cases
of uveal melanoma.
Which supports the hypotheses of
this paper.
Figure 1.

6. NESTIN EXPRESSION IN
PUM
In this figure A) has the red stain indicates nestin in the
neuroretina and could be used as a control to show that the
staining does work. B was the retinal pigment and stained
blue so was considered negative, C was primary uveal
melanoma and there indicates some nestin from looking at
it, D was the tissue section of PUM but attained less than
10% so was considered negative. E is normal melanocytes
which do not contain nestin so blue stain is what should be
expected.
Figure 2.

7. NESTIN EXPRESSION IN
PUM
A/B show 10-50% nestin expression
whereas C/D indicate greater than
50%.
Figure 3.

8. NESTIN EXPRESSION IN
MUM
21/26 of the MUM contained a high
proportion of nestin positive
tumour cells(A). B shows that 17/26
of MUM had closed loops. C shows
the 11% that were negative for
nestin positive cells. D shows that
nestin also stained regular
melanoma cells.
Figure 4.

9. NESTIN EXPRESSION IN PUM
COMPARED TO MUM
 Nestin expression in two representative matched PUM‐MUM. A, C, E and G are
H&E‐stained images. B, D, F and H show nestin expression. A high proportion of
nestin‐expressing tumour cells are seen in both (B) PUM1 and (D) MUM1 from the same
patient
Figure 5.

10. RESULTS FROM STUDY
• This paper shows a correlation between high levels of nestin and survival
time.
• There is also a link to the cell morphology and mitotic counts in relation
to nestin levels shown in figure 1.
• These numbers support the original hypotheses.
• Other papers state that nestin was higher in cutaneous melanoma
patients with stage IV disease compared to stage III/II patients (Fusi
et al. 2010). Which is similar to this study.
• In MUM, a higher percentage of nestin‐positive tumour cells combined
with poor prognostic markers in the PUM led to a shorter survival time
following the development of metastases.
• By the end of the study, four patients with MUM were alive while 21 had
died of their disease, and one patient was lost to follow‐up. The median
survival time following detection of metastatic disease was 12 months
(range 0.5–48).

11. CRITICAL APPRAISAL
• The number of PUM was good although the number of MUM could’ve
been greater although it being rare.
• The data gathered was sufficient and supportive of the hypothesis.
• More methods could have been used to show various things such as FISH
to identify any change in chromosome.
• Only 1 method of staining used (H&E)
• The paper is only small so the impact factor will be small
• The impact factor of this paper was 2, when compared to other papers
on this topic this is relatively low.
In a study on PRAME qPCR was used to analyse if it could be a successful
biomarker for metastases this study showed. (Field, et al 2016).