Work Activities

1. Selection of patients, communication with them (according to the requirements of the ethics committee), collection of material

Responsible persons: Doc. MUDr. Miroslav Korbeľ CSc, Doc. MUDr. Jozef Záhumenský, PhD.

The recruitment of patients (approximately 200) takes place during regular medical check-ups, and the patient are properly informed about the nature and significance of the project as well as the (not) risks of patient involvement in the project. Ethical aspects were evaluated by a local ethics committee. The patient receives the text "Patient Information" and "Informed Consent of Patient". At the same time, the "Relevant Patient Information Form" are completed by doctors. Samples are taken from the placenta of healthy women and the placenta of women with symptoms of preeclampsia (various stages and complications), after natural / in vitro fertilization. Samples are transported in appropriate solutions and subsequently submitted to histological, immunolocalization, genetic and proteomic analysis. Anonymous data from the "Relevant Patient Information Form" are provided for evaluation, statistical processing and interpretation of analysis results. The results are represented by the set of clinically characterized samples needed for further work activities.

2. Histological characterization of analyzed tissues, immunolocalization of syncytin-1 in preeclamptic and physiologically normal samples

Responsible persons: Prof. MUDr. Ľudovít Danihel, CSc, MUDr. Katarína Letkovská, PhD.

Samples taken from the placenta tissue are fixed in formalin (transported in this way) and embedded in paraffin wafers. Preparations is made from samples by a standard procedure and stained with hematoxylin and eosin to be evaluated by a light microscopy. Selected samples are processed by immunofluorescence and syncytin-1 expression is evaluated in a fluorescence microscope. Differences in syncytin-1 expression in placental tissue of healthy and preeclamptic patients are statistically processed.

3. Genomics and proteomics

Responsible persons: Doc. Ing. Helena Gbelcová, Ph.D, Mgr. Petra Priščáková, PhD.

Samples taken from the placenta tissues transported in the appropriate buffer are subjected to the genetic variability test of the ERVW-1 regulatory region by the direct sequencing method. Expression of the wild-type ERVW-1 gene and the ERVW-1 gene with the occurrence of the DNA variant are compared by RT-PCR (mRNA detection) and western blotting (determination of ERVW-1 gene product syncytin-1). These methods are used to determine the effect of a given DNA variant on changes in the amount of syncytin-1 whose abnormal amounts are described in preeclampsia. In case of real-time PCR, we use specific TaqMan probes. In Western blot, we use antibodies against both syncytin-1 subunits.

4. Statistical processing, evaluation, interpretation and dissemination of results

Responsible persons: RNDr. Katarína Sušienková, PhD.

Based on the selected parameters of the anamnesis of women involved in the project, we categorize them. The results obtained are evaluated using statistical methods to confirm the diagnostic significance of the identified DNA variants and their impact on ERVW-1 gene expression as well as on severity of the course of the disease.

5. Database

Variants


			VARIANT	rs112171707	rs150596337	-	rs555416193	rs149888808	-	rs1238265137	rs192755521	rs4727276	rs569899772	rs990071845	rs944910258	rs911934432	rs187902619	rs191798317	rs144621394
			LOCALIZATION WITH RESPECT TO ERWV-1	c.-2378C>A	c.-2372G>A	c.-2349_-2350insA	c.-2084C>T	c.-2048G>A	c.-1973A>T	c.-1972G>T	c.-1514T>C	c.-1284G>C	c.-1068C>T	c.-667A>G	c.-664A>G	c.-644A>G	c.-631T>C	c.-628C>A	c.-519C>T
			GENOMIC LOCALIZATION	92479532	92479526	92479503 - 92479502	92479238	92479202	92479127	92479126	92478668	92478438	92478222	92477821	92477818	92477798	92477785	92477782	92477673
SEQUENCE NUMBER	PE / FN	DIAGNOSIS	NUMBER OF MUTATIONS
1	FN		1									homozyg.
2	FN	gestačná hypertenzia	1															heterozyg.
3	FN		0
4	HD	PE, HELLP	0
5	HD	PE	2									heterozyg.							heterozyg.
6	FN		2		heterozyg.													heterozyg.
7	FN		1									heterozyg.
8	FN		1						heterozyg.
9	FN		0
10	HD	PE	2		heteroyg.														heterozyg.
11	HD	PE, HELLP	1	homozyg.
12	FN		0
13	FN		0
14	HD	PE	0
15	FN		0
16	HD	gestačná hypertenzia	1									heterozyg.
17	FN		1	heterozyg.
18	FN		1		heterozyg.
19	FN		0
20	HD	PE	0
21	FN		2	heterozyg.								heterozyg.
22	HD	IUGR	1	heterozyg.
23	FN		2							heterozyg.		hetrozyg.
24	HD	PE neskorá	1				heterozyg.
25	FN		1									heterozyg.
26	FN		1	heterozyg.
27	FN		0
28	FN		1									homozyg.
29	FN		1		heterozyg.
30	FN		1								heterozyg.
31	HD	getačná hypertenzia	1									heterozyg.
32	FN		0
33	HD	PE neskorá	0
34	FN		0
35	FN		0
36	FN		0
37	FN		1									heterozyg.
38	HD	PE, HELLP	0
39	FN		0
40	FN		0
41	FN		0
42	HD		1									heterozyg.
43	HD		0
44	HD		0
45	HD		1	heterozyg.
46	HD		1									heterozyg.
47	HD		0
48	HD		2	heterozyg.			heterozyg.
49	HD		1									heterozyg.
50	HD		0
51	HD		0
52	HD		1									homozyg.
53	HD		2	heterozyg.							heterozyg.
54	HD		0
55	HD		0
56	HD		1			homozyg.
57	HD		0
58	HD		0
59	HD		1									heterozyg.
60	HD		1									heterozyg.
61	HD		1									heterozyg.
62	HD		1									heterozyg.
63	HD		0
64	HD		1									heterozyg.
65	HD		2		heterozyg.							heterozyg.
66	HD		1									heterozyg.
67	HD		1									heterozyg.
68	HD		1									heterozyg.
69	HD		1									homozyg.
70	HD		0
71	HD		2	heterozyg.								heterozyg.
72	HD		1	heterozyg.
73	HD		0
74	HD		1									heterozyg.
75	HD		0
76	HD		2		heterozyg.							heterozyg.
77	HD		2				heterozyg.					heterozyg.
78	HD		0
79	HD		2	heterozyg.								heterozyg.
80	HD		1		heterozyg.
81	HD		0
82	HD		0
83	HD		2	heterozyg.								heterozyg.
84	HD		0
85	HD		0
86	HD		1									heterozyg.
87	HD		1									heterozyg.
88	HD		1									heterozyg.
89	HD		0
90	HD		1				heterozyg.
91	FN		1	heterozyg.
92	FN		0
93	FN		1	heterozyg.
94	FN		1										heterozyg.
95	FN		0
96	FN		1	heterozyg.
97	FN		1				heterozyg.
98	FN		1									heterozyg.
99	HD	PE	0
100	HD	PE	1									heterozyg.
101	HD	PE	1									heterozyg.
102	FN		0
103	HD	Gestačná hypertenzia, HELLP	0
104	HD	PE	0
105	HD	PE	1									heterozyg.
106	HD	Gestačná hypertenzia	1	heterozyg.
107	FN		1								heterozyg.
108	HD	PE (gestačná hypertenzia, IUGR)	1									heterozyg.
109	FN		0
110	FN		0
111	HD	PE	0
112	HD	PE	0
113	FN		0
114	FN		0
115	FN		0
116	FN		1									homozyg.
117	FN		1									heterozyg.
118	FN		0
119	FN		0
120	FN		0
121	FN		0
122	FN		1									heterozyg.
123	FN		0
124	HD	IUGR neskorá	0
125	FN		0
126	FN		0
127	FN		0
128	FN		1									homozyg.
129	FN		0
130	FN		1									heterozyg.
131	FN		1									heterozyg.
132	FN		1	heterozyg.
133	FN		0
134	FN		1									heterozyg.
135	FN		1									heterozyg.
136	FN		0
137	FN		0
138	FN		1									heterozyg.
139	FN		0
140	FN		1									heterozyg.
141	FN		1									heterozyg.
142	FN		0
143	FN		5	homozyg.										heterozyg.	heterozyg.	heterozyg.	heterozyg.
144	FN		0
145	FN		1									heterozyg.
146	FN		1									heterozyg.
147	FN		0
148	FN		1									heterozyg.
149	FN		1					heterozyg.
150	FN		1	heterozyg.
151	FN		1									homozyg.
152	HD	PE neskorá	0
153	FN		1									heterozyg.
154	FN		0
155	FN		1									heterozyg.
156	FN		1									heterozyg.

Variants vs QPCR


			VARIANT	rs112171707	rs150596337	-	rs555416193	rs149888808	-	rs1238265137	rs192755521	rs4727276	rs569899772	rs990071845	rs944910258	rs911934432	rs187902619	rs191798317	rs144621394	ERVW-1				ERV-FRD
			LOCALIZATION WITH RESPECT TO ERWV-1	c.-2378C>A	c.-2372G>A	c.-2349_-2350insA	c.-2084C>T	c.-2048G>A	c.-1973A>T	c.-1972G>T	c.-1514T>C	c.-1284G>C	c.-1068C>T	c.-667A>G	c.-664A>G	c.-644A>G	c.-631T>C	c.-628C>A	c.-519C>T	ΔCt (CtE1 - CtRP)		ΔCt (CtE1 - CtYW)		ΔCt (CtS2 - CtRP)		ΔCt (CtS2 - CtYW)
			GENOMIC LOCALIZATION	92479532	92479526	92479503 - 92479502	92479238	92479202	92479127	92479126	92478668	92478438	92478222	92477821	92477818	92477798	92477785	92477782	92477673	Δ Ct mean	Δ Ct SD	Δ Ct mean	Δ Ct SD	Δ Ct mean	Δ Ct SD	Δ Ct mean	Δ Ct SD
SEQUENCE NUMBER	PE / FN	DIAGNOSIS	NUMBER OF MUTATIONS
99	HD	PE	0																	1.4134	0.9377	0.6648	0.3267	4.5459	1.3504	3.9452	0.9177
100	HD	PE	1									heterozyg.								1.2182	0.8147	0.5470	0.4187	4.1141	0.8256	3.4429	0.4777
101	HD	PE	1									heterozyg.								0.1578	1.4387	0.4515	0.4607	4.6476	0.7603	4.9413	0.6329
102	FN		0																	4.3630	0.9971	0.1713	0.3717	7.9594	0.8321	3.7678	0.3474
103	HD	Gestačná hypertenzia, HELLP	0																	0.8059	1.2528	0.4466	0.2349	5.2498	1.7061	4.8905	0.9873
105	HD	PE	1									heterozyg.								0.4374	0.9333	-0.5488	1.3098	5.7495	0.8978	4.7634	0.6123
106	HD	Gestačná hypertenzia	1	heterozyg.																1.8004	0.6756	1.0843	0.3403	7.0754	0.5542	6.3593	0.4095
107	FN		1								heterozyg.									1.2149	0.8900	1.1736	0.6794	6.6219	0.7422	6.5805	0.6673
108	HD	PE (gestačná hypertenzia, IUGR)	1									heterozyg.								2.0150	1.1700	1.1079	0.4018	7.4130	0.9741	6.5059	0.3949
109	FN		0																	4.0869	1.1400	1.1964	0.9440	7.6719	0.7046	4.7814	0.1718
111	HD	PE	0																	0.4492	0.6274	0.1644	0.2314	6.7174	0.5311	6.4326	0.6552
112	HD	PE	0																	0.8764	0.6065	0.8677	0.1549	2.7593	0.6119	2.7506	0.2120
114	FN		0																	1.3747	0.8436	0.5877	0.5258	7.3188	0.7861	6.5317	0.3868
115	FN		0																	3.3568	0.8530	1.6060	0.5593	5.2832	1.1313	3.5324	0.3974
116	FN		1									homozyg.								2.5005	1.0524	1.9577	0.2815	3.9206	1.2756	3.3778	0.4341
117	FN		1									heterozyg.								3.3266	2.9804	2.1173	1.3554	7.0635	2.6684	5.8543	1.1802
118	FN		0																	2.1802	0.8667	1.5217	0.2835	5.6185	0.6514	4.9600	0.2185
119	FN		0																	4.0019	1.5069	0.8200	0.5375	7.6594	0.9787	4.4776	0.5071
120	FN		0																	4.7657	0.9239	1.6143	0.5265	8.6486	0.7929	5.4972	0.5318
121	FN		0																	3.7820	1.1105	1.4142	0.2580	8.1926	0.9483	5.8247	0.3440
122	FN		1									heterozyg.								3.5133	1.0103	0.6996	0.2596	8.1731	1.1359	5.3594	0.2436
123	FN		0																	3.6271	0.3983	1.5418	0.2447	5.7865	0.7087	3.7012	0.3792
124	HD	IUGR neskorá	0																	3.0764	0.7574	0.1766	0.1832	7.2688	0.8060	4.3691	0.2884
125	FN		0																	3.2427	1.1223	0.3470	0.9354	7.0136	1.0348	4.1179	0.8537
126	FN		0																	3.6387	0.7506	0.4176	0.4044	7.1477	0.6936	3.9266	0.3267
127	FN		0																	4.5577	2.3603	0.8295	1.1277	8.5424	2.3164	4.8141	0.8121
128	FN		1									homozyg.								0.3585	1.0915	0.2119	0.1944	4.4377	0.9090	4.2911	0.3743
133	FN		0																	1.7730	1.5821	-0.4190	0.4779	6.0423	1.4003	3.8502	0.3184
134	FN		1									heterozyg.								-0.6554	0.4183	0.0032	0.3352	3.6942	0.5198	4.3528	0.5541
135	FN		1									heterozyg.								2.8249	0.6738	1.1743	0.1176	5.9834	0.6641	4.3328	0.3804
136	FN		0																	2.9805	0.7812	0.9014	0.1208	6.3754	0.6906	4.2962	0.2599
137	FN		0																	4.5014	1.8402	2.2747	1.9852	6.5904	0.5976	4.3637	0.4882
138	FN		1									heterozyg.								2.9474	1.4449	1.6593	0.2144	6.3402	1.3965	5.0521	0.3268
139	FN		0																	4.8607	0.7941	1.2152	0.4250	7.5853	0.7655	3.9398	0.3466
140	FN		1									heterozyg.								3.9624	0.3158	1.5593	0.3581	6.4141	0.6947	4.0110	0.4944
141	FN		1									heterozyg.								3.8290	0.7537	1.1973	0.4245	5.7411	0.8502	3.1094	0.3319
142	FN		0																	3.0324	0.7408	1.3878	0.2497	5.6662	0.6276	4.0216	0.2558
143	FN		5	homozyg.										heterozyg.	heterozyg.	heterozyg.	heterozyg.			5.2983	0.7600	1.3591	0.2990	5.8489	0.7961	3.9885	0.3122
144	FN		0																	5.6677	1.4458	2.3632	1.0129	5.0556	3.0892	1.7511	3.4425
145	FN		1									heterozyg.								3.5824	0.9570	1.7800	0.2465	6.4006	0.6330	4.5982	0.3749
146	FN		1									heterozyg.								5.2655	0.7142	2.2089	0.2623	8.1686	0.4093	5.1121	0.3720
147	FN		0																	4.3029	0.2671	1.4371	0.1796	7.5895	0.2340	4.7237	0.1347
148	FN		1									heterozyg.								5.3333	0.8927	1.9071	0.3284	8.0407	0.6341	4.6144	0.2032
149	FN		1					heterozyg.												5.4401	2.1752	2.0332	1.0555	8.9771	1.9538	5.5702	1.0799
150	FN		1	heterozyg.																4.4193	0.8273	1.6257	0.2582	7.6462	0.8000	4.8526	0.5104
151	FN		1									homozyg.								3.0615	0.7218	1.5017	0.1541	5.7227	0.7820	4.1628	0.2797
152	HD	PE neskorá	0																	1.7481	1.1223	1.1972	0.3068	3.5044	0.9349	2.9535	0.2982
153	FN		1									heterozyg.								4.3585	1.1463	1.7712	0.4883	6.1453	0.8530	3.5579	0.4931

SYNCYTIN-1 as a new marker for pre-eclampsia, the second leading cause of death in pregnant women

Work activities

1. Selection of patients, communication with them (according to the requirements of the ethics committee), collection of material

2. Histological characterization of analyzed tissues, immunolocalization of syncytin-1 in preeclamptic and physiologically normal samples

3. Genomics and proteomics

4. Statistical processing, evaluation, interpretation and dissemination of results

5. Database

Variants

Variants vs QPCR

Project supported by the Ministry of Health of the Slovak Republic (MZSR 2018/40-LFUK-14)

SYNCYTIN-1 as a new marker for pre-eclampsia, the second leading cause of death in pregnant women

Work activities

1. Selection of patients, communication with them (according to the requirements of the ethics committee), collection of material

2. Histological characterization of analyzed tissues, immunolocalization of syncytin-1 in preeclamptic and physiologically normal samples

3. Genomics and proteomics

4. Statistical processing, evaluation, interpretation and dissemination of results

5. Database

Variants

Variants vs QPCR

Project supported by the Ministry of Health of the Slovak Republic (MZSR 2018/40-LFUK-14)

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