Introduction

I expect you to recognize basic tissue types and neoplasms, based on clinical and pathologic data. Images may be a part of that information, but will not be the only clues provided (unless there is a defining pathologic feature that gives you the diagnosis immediately - that I would expect you to know). Remember: my primary goal is not to have you to regurgitate simple facts back to me on the exam.  Rather, I want you to be able to synthesize, analyze, and USE the information I have given you.

Again, my job is to teach you to think clinically.  I will ask straightforward questions, designed to test whether you know (very) basic morphology and elements that go into a pathologic diagnosis, as well as being able to reason through a differential diagnosis to reach a final answer, taking into account clinical presentation, physical exam, labs, and any secondary screening tests, as necessary. 

As for suggested study topics, I want you to know how to differentiate between clinical disease states; the difference between benign and malignant neoplasms (as well as borderline tumors, if relevant) IN GENERAL; and the clinicopathologic differences between tumor categories that are specific enough to narrow your differential.

Don’t overthink it. I will ask no trick questions. You’re second-years, not pathology residents, and I am fully cognizant of that fact.  All I want, is for you to be able to ask the right questions to get to a diagnosis, and understand what that diagnosis means in terms of clinical importance, prognosis, epidemiology, etc.

Hematology and Lymph Lectures

Hematopoietic Neoplasms - For Your Interest

Normally, when people ask me what areas of a topic they should study, I get frustrated: you can’t be an effective physician if you only know the part of medical knowledge that’s in BOLD PRINT.

HOWEVER, since hematopoietic neoplasms (especially lymphomas) are fairly unique in that they often look exactly the same, morphology is often of limited value, and these entities have to be differentiated by immunophenotype and molecular studies.

Therefore, I would suggest you focus your attention on the information I have given you in the following hierarchy of importance (for each entity):

  1. Similarities to other neoplasms at the same level of maturation (in order to develop a differential diagnosis), including immunophenotype and morphology 
  2. Differences between other neoplasms at the same level of maturation, including immunophenotype and any molecular/cytogenetic markers that are specific for that disease entity (in order to narrow the differential diagnosis)
  3. Unique features (if any) that separate any individual entities from all others, including immunophenotype, morphology, molecular, treatment, and specific diagnostic tests that are only seen in that specific disease entity
  4. Clinical features, such as prognostic markers, aggressiveness, high risk/prevalence patient populations
  5. Pathologic features (immunophenotype and morphology) of the NORMAL correlate to each entity (i.e. the features of the normal lymphocyte at the same level of maturation), unless it differs from the neoplasm in a very specific way that can be exploited in diagnosis or treatment

You do not have to memorize all the pictures. They are there for reference.  I expect you to recognize basic tissue types and neoplasms, based on clinical and pathologic data.  Images may be a part of that information, but will not be the only clues provided (unless there is a defining pathologic feature that gives you the diagnosis immediately - that I would expect you to know). Remember: my primary goal is not to have you to regurgitate simple facts back to me on the exam.  Rather, I want you to be able to synthesize, analyze, and USE the information I have given you.

Again, my job is to teach you to think clinically.  I will ask straightforward questions, designed to test whether you know (very) basic morphology and elements that go into a pathologic diagnosis, as well as being able to reason through a differential diagnosis to reach a final answer, taking into account clinical presentation, physical exam, labs, and any secondary screening tests, as necessary. 

As for suggested study topics, I want you to know how to differentiate between clinical disease states; the difference between benign and malignant neoplasms (as well as borderline tumors, if relevant) IN GENERAL; and the clinicopathologic differences between tumor categories that are specific enough to narrow your differential.

Don’t overthink it. I will ask no trick questions. You’re second-years, not pathology residents, and I am fully cognizant of that fact.  All I want, is for you to be able to ask the right questions to get to a diagnosis, and understand what that diagnosis means in terms of clinical importance, prognosis, epidemiology, etc.

Study Stratgeies

I have noticed some trends in the questions I've received on these lectures over the years, so I have some general suggestions:

  1. Try to summarize/present each disease entity to either one of the tutors, or one of your colleagues.  Putting the material in your own words will better anchor the information, because it will be more consistent with how YOU think, rather than how I think.  Also, if the person you’re presenting to is knowledgeable about the subject, they might be able to point out gaps in your knowledge, which can then be remedied.
  2. For the immunophenotype markers, focus on the ones that were highlighted as clinching the diagnosis of specific entities.  If you get down to two or three possibilities, look for clues in the clinical scenario that might knock out one of the related entities, and narrow your differential.
  3. Talk to Student Services about study habits that have proven successful for previous students.  They may have ideas you haven’t thought of.
  4. Remember that my areas of specialization are hematopathology and molecular pathology.  So it’s probably a safe bet that cytogenetic and molecular features that either define a diagnosis, or have a major impact on prognosis, are going to be high yield topics.
  5. Download and review the lectures as powerpoint files, rather than PDF’s.   I have extensive annotations in the Notes section of each slide that I follow in lecture to make sure I hit all the points that I want to emphasize.
  6. Dr. Lee's lecture block will be approaching the same disease entities as my lectures, but from a more clinical viewpoint.  In addition to the new material, use them as a refresher tool in conjunction to the earlier presentation, to check whether the material comes easier the second time.  Reinforcement and rephrasing are key to getting the information to stick in your mind.
  7. If you have any specific areas of difficulty, feel free to reach out to the lecturer (me, for the most part) for clarification.  I will respond ASAP to emails, and I have no problem setting up one-on-one Zoom conferences, if you can’t make my open office hours.  That IS my job, after all.  If you fail, I’ve failed.

I don’t expect you to be pathologists.  I only want you to recognize the basic uses of the various stains I touched on in class, as well as morphologic findings which are SPECIFICALLY linked to a particular organism or condition.  If it's not uniquely associated with a particular organism or disease entity, then I don't consider it particularly important at this stage in your training.

In general, on exams I will try to give you multiple clues in a patient scenario, whether clinical, laboratory, or morphologic, so that you can come to the right answer in multiple ways.  The importance is to narrow your differential to get a diagnosis, and then know enough about that diagnosis to be able to answer the question.  I mostly want you to learn how to think through a problem to reach a conclusion, and less so necessarily to just memorize images.

Specific Expectations

You need to be able to recognize (histologically) the difference between follicular (FL), diffuse (DLBCL, LBL, ALCL, extranodal NK/T), and starry sky architectural patterns (Burkitt, but also may be seen in LBL and DLBCL), and know the lymphomas associated with them. 

You need to be able to recognize (cytologically) neoplasms with distinctive cell morphology: classic Hodgkin lymphoma, anaplastic large cell lymphoma (ALCL), adult T cell leukemia/lymphoma (ATLL), acute promyelocytic leukemia (APL), AML with inv16, etc.

You need to know the distinctive cytogenetic abnormalities that underly various neoplasms (e.g. CML has a 9;22 translocation), as well as specific molecular lesions that define hematopoietic disease entities (eg. t(9;22) in CML = BCR-ABL1 fusion).  

Lastly, you need to know the phenotypic markers that distinguish specific neoplasms from each other, and the context in which they are diagnostically important:  eg. naive B cell tumors (CLL and MCL) are both positive for CD20 and CD5.  However, MCL is also positive for BCL1, while CLL is not (CLL is also positive for LEF1, unlike MCL). 

Immunophenotype (CD Markers)

In the broadest sense, B lineage cells have three stages of development: (blast, mature lymphocyte, and plasma cell), and the mature B lymphocyte subset has three major steps in maturation (Pre-GC, GC, and Post-GC).  T cells can be said to have two stages of development (blasts to lymphocyte), and mature T lymphocytes go through three (main) stages of development: CD4/CD8 double negative, CD4/CD8 double positive, and then single positive (CD4 or CD8). 

  • A 'pan' marker is positive at all stages of development.  B lineage cells are positive for CD19, whether blasts, lymphocytes, or plasma cells.  Therefore CD19 is a pan-B marker.
  • Mature B cell markers are only positive in the lymphocyte stage.  CD20 is a mature B lymphocyte marker.  
  • CD2 is positive in all T lineage cells, whether blast or lymphocyte.  Therefore CD2 is a pan-T marker.  
  • CD3 is a mature marker, because it is only positive in mature T cells (Mostly. CD3 can actually be positive in late blasts and has partial expression in NK cells).
  • Hematopoietic stem cells (HSC), which are not committed to B or T lineage, do not express CD2, CD3, CD19, or CD20, but are instead positive for CD34, a stem cell marker.  
  • TdT is a blast marker, for both B and T lineages. 
  • B blasts are positive for CD34, TdT, and CD19. Mature B cells lose CD34 and TdT, but gain CD20.
  • T blasts are positive for CD34, TdT, and CD2.  Mature T cells are positive for CD2 and CD3.


 

The Ontogeny Model

A LOT of people have expressed confusion about CD markers and which ones I expect you to know.  I realize it can be overwhelming, but it’s the best way to tell these tumors apart (and this is how we do it in clinical practice).  More information on each marker is included in the Appendices, for reference.  This additional material is for your information (FYI) only.  You will not be tested on the Appendix material.

What I want you to understand from all this, is that markers are clustered by lineage, stage of development, stage of maturation, etc.  In this way, you can tell one lymphocyte from another, even if they look the same under the microscope.  By analogy, you can tell the lymphomas apart by the phenotype of the cells from which they are derived (the 'Ontogeny' model).

These are the markers I want you to know.  I would suggest you break down the different markers for each disease by the unique features of their corresponding normal counterpart:

 

Normal:

  • Hematopoietic Stem Cell: CD34+
  • B-blast: CD19+, CD10+, CD34+, TdT+, sIg-negative
  • Naïve B cell: CD19+, CD20+, BCL2+
  • Germinal center B cell: CD19+, CD20+, CD10+, BCL6+, BCL2-
  • Activated B cell: CD19+, CD20+, MUM1+, surface Ig+
  • Plasma cell: CD138, CD19+, CD20-, MUM1+, cytoplasmic Ig+

 

  • T-blast/thymocyte: CD2+, CD34+, TdT+, CD3 +/-,  (CD4-CD8- or CD4+CD8+)
  • Mature helper T cell: CD3+, CD4+
  • Mature follicular helper T cells:  CD3+, CD4+, CD10+, BCL6 +/-
  • Mature cytotoxic/regulatory T cell: CD3+, CD8+, cytotoxic markers (TIA1+, perforin+, granzyme B+)

 

B cell lymphomas

  • Follicular lymphoma (derived from germinal center B cells): CD19, CD20, CD10, BCL6, BCL2
  • Marginal zone lymphoma (derived from activated, post-germinal center B cells): CD19, CD20, MUM1
  • Mantle cell lymphoma (derived from naïve or memory B cells): CD19, CD20, CD5, BCL1+
  • CLL/SLL (derived from naïve B cells): CD19, CD20+ dim, CD5, BCL1-, CD23+, LEF1+, sIg+ dim
  • Burkitt lymphoma (derived from germinal center B cells with a MYC translocation): CD19, CD20, CD10, BCL-6, MYC+, Ki-67 ~100%
  • Diffuse large B cell lymphoma:  CD19, CD20, Ki-67 40-80% (may show either Germinal center or Activated B cell markers)
  • B-Lymphoblastic leukemia/lymphoma (derived from B blast): CD19, CD34, TdT

 

Oddballs: B cell lymphomas without a well-defined normal counterpart

  • Classic Hodgkin lymphoma: CD30+, CD15+/-, CD45-, CD20-, CD3-, Reed-Sternberg cells and variants (e.g. lacunar cells in NS CHL)
  • Nodular lymphocyte predominant Hodgkin Lymphoma: CD19, CD20, CD45, ‘popcorn’ cells
  • Hairy Cell Leukemia:  BRAF V600E, CD20, CD25, CD103, CD11c, BCL1+ in 50% of cases, TRAP+ (obsolete)

 

T cell lymphomas

  • Angioimmunoblastic T cell Lymphoma (derived from T follicular helper cells): CD3, CD10, CD4 (associated reactive vascular proliferation and reactive clonal expansion of EBV+ B cells à polyclonal gammopathy)
  • Adult T cell leukemia/lymphoma (derived from T helper cells): CD3, CD4, HTLV1+
  • Anaplastic large cell lymphoma (derived from cytotoxic T cells): CD30, ALK1 +/-, CD3+/-, cytotoxic markers (TIA1+, perforin+, granzyme B+)
  • Enteropathy associated T cell lymphoma (derived from intraepithelial lymphocytes, CD4-/CD8-): CD3+, CD4-, CD8-, celiac disease
  • T-lymphoblastic leukemia/lymphoma (derived from T blasts): CD3, CD34, TdT, CD4+/CD8+ or CD4-/CD8-, typically presents as an adolescent male with a mediastinal mass

 

Non-lymphoid hematopoietic malignancies:

  • Mastcytosis (derived from mast cells, CD25+): CD117, CD25, tryptase+, typically KIT (CD117) amino acid 816 is mutated from glutamate to a hydrophobic side group, most commonly valine (KITD816V)
  • Blastic plasmacytoid dendritic cell neoplasm (derived from plasmacytoid dendritic cells, CD123+): CD3+/-, CD4, CD56, CD123
  • Extranodal NK/T cell lymphoma (derived from NK or NKT cells, CD56+, cytoplasmic CD3+, surface CD3 +/-): cytoplasmic CD3+, surface CD3+/-, CD56+, EBV+ in ~100% of tumor cells
  • Acute myeloid leukemia: CD34+, TdT+/-, myeloperoxidase+.  Other myeloid markers are of limited utility compared to morphology and genetic/molecular studies.

Final Thoughts

On a philosophical note, memorization may be required to master the Science of medicine, and you have your entire careers to learn it.  There's a lot to learn, and you'll need it for board exams like the COMLEX, but that won't make you doctors.

Analysis is the Art of medicine, and that the most important preparation for clinical practice next year, when you hit your clerkships.  Learn to think through a problem, from the clinical context, history, and physical evidence, to get to a differential diagnosis.  Then determine what more information you need to get to a final diagnosis, and how to acquire it.  Once you identify a diagnosis, that will inform your treatment decisions, which is the ultimate goal of everything you have done up to this point. 

I do want you to learn the science, because that's the basis for the knowledge you will gain throughout the rest of your professional lives.

But if I can help you learn how to think, then I have achieved my highest goals. Good Luck.

 

 

Respectfully,

Dr. Ahsanuddin

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