fa شناسایی 5 جهش جدید در ژن گلیکوپروتئین Ibα پلاکت در بیماران برنارد- سولیر ایران خون و انکولوژی Hematology & oncology پژوهشي Research <h2 style="MARGIN: 0cm 0cm 0pt DIRECTION: rtl" dir="rtl"><p><span></span></p><p><span>    زمینه وهدف: سندرم برنارد سولیر بیماری خونریزی دهنده ارثی است که به دلیل نقایص مولکولی</span><span dir="ltr"></span><span dir="ltr" lang="AR-SA"><span dir="ltr"></span><font face="Times New Roman">�</font></span><span style="FONT-FAMILY: " lang="FA">کمپلکس </span><font face="Times New Roman"><span dir="ltr">GP Ib-IX-V</span><span dir="rtl"></span></font><span style="FONT-FAMILY: "><span dir="rtl"></span> </span><span dir="ltr"></span><span dir="ltr"><span dir="ltr"></span><font face="Times New Roman">(GlycoProtein)</font></span><span dir="rtl"></span><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span> پلاکتی که</span><span style="FONT-FAMILY: " lang="FA"> از چهار زنجیره </span><font face="Times New Roman"><span style="mso-bidi-language: FA" dir="ltr">GP </span><span dir="ltr">Ibα</span><span style="mso-bidi-language: FA" dir="ltr">, GP </span><span dir="ltr">Ibβ</span><span style="mso-bidi-language: FA" dir="ltr">, GP IX, GP V</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="FA"><span dir="rtl"></span> تشکیل شده است، </span><span style="FONT-FAMILY: " lang="AR-SA">اتفاق می‌افتد.</span><span style="FONT-FAMILY: " lang="AR-SA">�</span><font face="Times New Roman"><span dir="ltr">GP Ibα</span><span dir="rtl"></span></font><span style="FONT-FAMILY: "><span dir="rtl"></span> <span lang="AR-SA"><font face="B Yagut">بزرگترین زنجیره این کمپلکس بوده و مسئول اتصال به لیگاند می‌باشد</font></span></span><span style="FONT-FAMILY: " lang="FA">.</span><span style="FONT-FAMILY: " lang="AR-SA"> بیشتر</span><span style="FONT-FAMILY: " lang="FA"> جهش</span><span style="FONT-FAMILY: " lang="AR-SA">‌های شناسایی شده</span><span style="FONT-FAMILY: " lang="FA"> در ارتباط با سندرم برناردسولیر</span><span style="FONT-FAMILY: " lang="AR-SA"> در </span><font face="Times New Roman"><span dir="ltr">GP Ibα</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span> بوده است. هدف از انجام این مطالعه بررسی و شناسائی <font face="B Yagut"><span style="mso-spacerun: yes"> </span>نقایص مولکولی ژن</font></span><span style="FONT-FAMILY: " lang="AR-SA">�</span><span dir="ltr"></span><span dir="ltr"><span dir="ltr"></span><font face="Times New Roman"><span style="mso-spacerun: yes"> </span>GP Ibα</font></span><span style="FONT-FAMILY: " lang="AR-SA">در بیماران برنارد سولیر ایرانی می‌باشد. </span><span style="FONT-FAMILY: " lang="FA"></span><h4 style="MARGIN: 0cm 0cm 0pt" dir="rtl"><span>روش‌ کار: این مطالعه به روش بررسی بیماران (</span><font face="Times New Roman"><span style="LETTER-SPACING: -0.1pt" dir="ltr">Case series</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="FA"><span dir="rtl"></span>) انجام شد و </span><span style="FONT-FAMILY: " lang="AR-SA">با استفاده ازبانک اطلاعاتی بیماری‌های خونریزی دهنده درمانگاه جامع هموفیلی ایران <font face="B Yagut"><span style="mso-spacerun: yes"> </span>12 بیمار</font></span><span style="FONT-FAMILY: " lang="FA"> شناسایی و</span><span style="FONT-FAMILY: " lang="FA">�</span><span style="FONT-FAMILY: " lang="AR-SA">مورد مطالعه قرار گرفتند. تشخیص بیماری بر پایه مطالعات فنوتیپی شامل شمارش پلاکت، بررسی لام خون محیطی و عدم پاسخدهی به آگونیست ریستوستین صورت گرفته بود. </span><span style="LETTER-SPACING: -0.1pt" dir="ltr"><font face="Times New Roman">NA</font></span><span style="FONT-FAMILY: " lang="AR-SA">D از ستونهای خونی<font face="B Yagut"><span style="mso-spacerun: yes">  </span>بیماران و افراد وابسته جدا سازی </font></span><span style="FONT-FAMILY: " lang="FA">شد. </span><span style="FONT-FAMILY: " lang="AR-SA">نواحی کد کننده ژن </span><font face="Times New Roman"><span style="LETTER-SPACING: -0.1pt" dir="ltr">GP Ibα</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span> به 5 قطعه هم پوشان تقسیم بندی و تکثیر انجام شد. ازروش </span><font face="Times New Roman"><span style="LETTER-SPACING: -0.1pt" dir="ltr">CSGE</span><span dir="rtl"></span></font><span style="FONT-FAMILY: "><span dir="rtl"></span> </span><span dir="ltr"></span><span style="LETTER-SPACING: -0.1pt" dir="ltr"><span dir="ltr"></span><font face="Times New Roman">(Conformation Sensitive Gel Electrophoresis)</font></span><span dir="rtl"></span><span style="FONT-FAMILY: "><span dir="rtl"></span> <span lang="AR-SA"><font face="B Yagut">استفاده گردید <span style="mso-spacerun: yes"> </span>و نهایتا <span style="mso-spacerun: yes"> </span>توالی‌های دارای هترودوبلکس در ژل </font></span></span><font face="Times New Roman"><span style="LETTER-SPACING: -0.1pt" dir="ltr">CSGE</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span> تعیین توالی شدند. </span><span style="FONT-FAMILY: " lang="FA"><p></p></span></h4><h4 style="MARGIN: 0cm 0cm 0pt" dir="rtl"><span>یافته‌ها: </span><span style="FONT-FAMILY: " lang="AR-SA">5 </span><span style="FONT-FAMILY: " lang="FA">جهش جدید شناسایی شد: حذف</span><font face="Times New Roman"><span dir="ltr">ACCGGCT </span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span><font face="B Yagut"><span style="mso-spacerun: yes"> </span>و جایگزینی </font></span><font face="Times New Roman"><span dir="ltr">GGA</span><span dir="rtl"></span></font><span style="FONT-FAMILY: "><span dir="rtl"></span> </span><span style="FONT-FAMILY: " lang="AR-SA">بین نوکلئوتید‌های 425-419، </span><span style="FONT-FAMILY: " lang="FA">جهش</span><span style="FONT-FAMILY: " lang="AR-SA">‌های نقطه‌ای </span><font face="Times New Roman"><span dir="ltr">C</span><span dir="rtl"></span></font>_{<span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span>709</span>}<font face="Times New Roman"><span dir="ltr">T</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span>،</span><font face="Times New Roman"><span dir="ltr">A </span><span dir="rtl"></span></font>_{<span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span>710</span>}<font face="Times New Roman"><span dir="ltr">G</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span>،</span><font face="Times New Roman"><span dir="ltr">T </span><span dir="rtl"></span></font>_{<span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span>1759</span>}<font face="Times New Roman"><span dir="ltr">C</span><span dir="rtl"></span></font><span style="FONT-FAMILY: " lang="AR-SA"><span dir="rtl"></span> و حذف 20 نوکلئوتید در موقعیت 1819-1800. موتاسیون‌های شناسایی شده به بانک جهانی ژن فرستاده شد و ثبت گردید برای هر کدام از موتاسیون‌ها </span><span dir="ltr"></span><span dir="ltr"><span dir="ltr"></span><font face="Times New Roman">(RFLP</font></span><font face="Times New Roman"><span style="mso-bidi-language: FA" dir="ltr">) Restriction Fragment Length Polymorphism</span><span dir="rtl"></span></font><span style="FONT-FAMILY: "><span dir="rtl"></span> <span lang="FA"><font face="B Yagut">طراحی وانجام شد.</font></span></span><span style="FONT-FAMILY: " lang="AR-SA"><p></p></span></h4><span dir="rtl" lang="AR-SA"><strong>نتیجه‌گیری</strong></span><span><strong>: </strong>در این مطالعه نقایص مولکولی ژن زنجیره </span><span>GP Ibα</span><span dir="rtl"></span><span dir="rtl"><span dir="rtl"></span> <span lang="AR-SA"><font size="3" face="B Yagut">پلاکتها در بیماران </font></span></span><span dir="ltr"></span><span><span dir="ltr"></span><font face="Times New Roman"><span style="mso-spacerun: yes"><font size="3"> </font></span>BSS </font></span><span dir="rtl" lang="AR-SA">مورد مطالعه قرار گرفت و 5 موتاسیون جدید که<b>�</b>در سایر مطالعات شناسایی نشده بود، شناسایی گردید</span><span dir="rtl" lang="FA">. </span></p></h2><p></p> <p style="MARGIN: 0cm 0cm 0pt" class="MatnLatin"><span><font face="Verdana"><strong></strong></font></span></p><p style="MARGIN: 0cm 0cm 0pt" class="MatnLatin"><span><font face="Verdana"><strong>    Background & Aim: </strong>Bernard-Soulier syndrome (B.S.S) is a rare hereditary bleeding disorder due to molecular defects of platelet GPIb–IX–V. The GPIb-IX-V complex is composed of four chains of GPIbα, GPIbβ, GPIX and GPV.<span style="mso-spacerun: yes">  </span>The largest chain of this complex is GPIbα and is responsible for binding to ligand and most of identified mutations belong to this glycoprotein.<span style="mso-spacerun: yes">  </span>The aim of<span style="mso-spacerun: yes">  </span>this<span style="mso-spacerun: yes">  </span>study was to identify the molecular defects of GPIbα gene in Iranian Bernard-Soulier<span style="mso-spacerun: yes">  </span>patients. </font></span><p></p><p style="MARGIN: 0cm 0cm 0pt" class="MatnLatin"><font face="Verdana"><b><span style="mso-bidi-language: FA" lang="EN">Patients and Method: </span></b><span style="mso-bidi-language: FA" lang="EN">Twelve Bernard-Soulier patients were selected from data base of bleeding disorders </span><span lang="EN">in </span><span style="mso-bidi-font-family: Calibri mso-bidi-language: FA" lang="EN">Comprehensive Clinic of Iranian Hemophilia Center. </span><span style="mso-bidi-language: FA" lang="EN">Diagnostic criteria for B.S.S were based on phenotypic analysis such as platelet counts, inspection of peripheral blood smear and lack of response to restocetin agonist. Genomic DNA was isolated from blood leukocytes of the patients and their parents. The entire amino acid coding region in exon 2 from GPIbα was divided into five overlapping fragments and PCR amplification was done. Finally, sequence analysis of the coding regions that contain DNA heteroduplexes in CSGE gels was performed.</span></font><p></p><p style="MARGIN: 0cm 0cm 0pt" class="MatnLatin"><font face="Verdana"><b><span style="mso-bidi-language: FA" lang="EN">Results: </span></b><span style="mso-bidi-language: FA" lang="EN">Sequence analysis revealed five novel mutations in GPIbα. The mutations include ACCGGCT deletion, GGA insertion in 419-425 position, missense mutations in T709C, G710A and C1759T, and the deletion of 20 nucleotides in 1800-1819 position. All five novel mutations were registered in International Gene Bank and for each mutation<span style="mso-spacerun: yes">  </span>RFLP (restriction fragment length polymorphism) design was created. </span><span style="mso-ansi-language: EN-US mso-bidi-language: FA"></span></font><p></p><p style="MARGIN: 0cm 0cm 0pt" class="MatnLatin"><font face="Verdana"><b><span style="mso-bidi-language: FA" lang="EN">Conclusion: </span></b><span style="mso-bidi-language: FA" lang="EN">In the present study, the molecular defects of GPIbα gene was investigated and five novel mutations were identified among Iranian BSS patients.</span></font><p></p></p><p></p></p><p></p></p><p></p></p><p></p> کلیدواژه ها: 1- سندرم برنارد-سولیر 2-گلیکوپروتئین Ibα 3- تعیین توالی 4- جهش Key Words: 1) Bernard-Soulier Syndrome 2) Glycoprotein Ibα 3) Sequence Analysis 4) Mutation 58 66 http://rjms.iums.ac.ir/browse.php?a_code=A-10-1-970&slc_lang=fa&sid=1 S.A Taghavi سیداسداله تقوی 3900319475328460016473 3900319475328460016473 No A Kazemi احمد کاظمی 3900319475328460016474 3900319475328460016474 Yes GH Rastegar Lari قاسم رستگار لاری 3900319475328460016475 3900319475328460016475 No F Ala فریدون علا 3900319475328460016476 3900319475328460016476 No Comprehensive Clinic of Iranian Hemophilia center درمانگاه جامع هموفیلی ایران، M Rasoolzadegan مریم رسول زادگان 3900319475328460016477 3900319475328460016477 No Comprehensive Clinic of Iranian Hemophilia center درمانگاه جامع هموفیلی ایران،